Atosfrcrepnec [email protected];[email protected]) ([email protected]; correspondence for *Authors Australia. 4072, QLD, ui,QD 02 Australia. 4072, QLD, Lucia, ai aiypoen n h sebyo of Kovtun assembly Oleksiy the and proteins family Cavin COMMENTARY ß ta. 92.Floigti,toohrhmlge fCAV1 specific muscle Rothberg of the 1 homologues and 1992; only 1996) other al., al., et two was (Scherer et this, CAV2 (CAV1) found, Following (Kurzchalia were 1992). -1 later al., years et component 1955), Yamada, 40 protein 1953; identified (Palade, first 1950s the the in (EM) 2013). microscopy Pozo, del a and Parton supplying in molecules (reviewed as surface – signalling cell well the membrane at of mechanoprotection as and mechanosensing plasma trafficking, for regulation reservoir membrane intracellular a the for enzymes, revealed of platform and including now organisation a – have caveolae and providing for studies turnover functions specialised of of lipid highly array cellular number diverse of large remarkably uneven suggestive A cells an profiles functions. muscle tissue- expression display distinct have specific and proteins generally caveola-associated and endothelial Caveolae distribution, adipocytes, 1A). cell many (Fig. in including membrane plasma the of 50– feature types, main of a invaginations are that bulb-shaped nm are 60 caves’) ‘little (or Caveolae Introduction Electron crystallography coil, X-ray Coiled microscopy, Cavin, Caveolin, Caveolae, WORDS: by KEY stress membrane meta- plasma protein–lipid to a cavins. of respond generating release can caveolae, and that low-affinity of structure formation coincident, stable caveola protein–protein the which for allow in interactions controlled important model a be with electrostatically propose interactions may identifying We that as function. well phospholipids a as provided membrane oligomerisation, have basis distinct their molecular labs the for and several stoichiometry form in cavin from of cell understanding Studies the better to into stimuli. released associate to be hetero-oligomerisation can that response In which members caveolae, and assembly. on family coat subcomplexes homo- cavin cavin four into structural through are insights highlighting there formation, new mammals, caveola provide in that cavins of studies role data the recent summarise on we peripheral Commentary, of this of regulators In family biogenesis. and caveola components cavin coat the past essential of as by the proteins emergence membrane However, the driven seen . has primarily be called decade to proteins formation in considered membrane whose generally membrane integral were plasma invaginations they the recently, membrane of Until feature cells. abundant many an are Caveolae ABSTRACT h nvriyo uesad nttt o oeua isine rsaeSt Brisbane Bioscience, Molecular for Institute Queensland, of University The 05 ulse yTeCmayo ilgssLd|Junlo elSine(05 2,16–28doi:10.1242/jcs.167866 1269–1278 128, (2015) Science Cell of Journal | Ltd Biologists of Company The by Published 2015. lhuhcvoa eeoiial bevdb electron by observed originally were caveolae Although 1 ia .Tillu A. Vikas , 2 etefrMcocp n iraayi,S.Lucia, St. Microanalysis, and Microscopy for Centre 1 ihlsAriotti Nicholas , 1 oetG Parton G. Robert , o h omto falcvoa,Cvn SP) Cavin3 (SDPR), Cavin2 (MURC). caveolae, Cavin4 all isoform muscle-specific There essential of the is 2001). and formation a CAV1, (PRKCDBP), al., displays like the et which and, (PTRF), Vinten profile for Liu Cavin1 expression 2005; 2008; mammals, broad al., al., in relatively et et 2004; cavins Hill four Vinten 2009; al., are al., 2008; et et al., Hansen (Aboulaich et 2009; of al., architecture understanding et our and Bastiani of proteins, assembly revision essential of a family and caveolar demanding cavin abundant defined, the 1985; been second caveolae, has a of al., decade, component only past structural the et the to caveolins. in time, ascribed (Peters membrane-embedded However, initially the a same was coating components, which protein possess the known 1992), spiral al., caveolae Around et or that Rothberg 1995). striated revealed techniques Parton, distinctive EM and specialised (Way CAV3 03 hese l,21) nti omnay eeaiethese examine we Commentary, this Pozo, in del 2014), and al., Parton (Bastiani et 2010; caveolae Shvets Sessa, 2013; of and biology Chidlow 2014; 2010; the Parton, al., reviews on and comprehensive et updates are Kovtun timely there provide 2013; Although that al., 2013). et EM al., association by Hansen et X-ray oligomers Ludwig 2014; cavin first cavin al., isolated et universal the of (Gambin visualisation the coat, the and of caveolar domain and the structure single-molecule of crystallographic biochemical, analysis Recent on including 2013). light ultrastructural shed to Pozo, processes, beginning are del that these data muscle and new and provided Parton lipid have studies and in in cancer, (reviewed role including their diseases dystrophies signalling, in defining cell role of and their the terms and biogenesis in of caveolar interest disassembly remodelling, molecular major and membrane of the assembly is Understanding to coat the similarity cavin proteins. govern structural coat that clear no mechanisms known show other which families of of any two mechanism just both by cooperative protein, mediated highly of primarily (Ludwig is a components that suggests formation coat This caveola caveolar 2013). major al., the et 1998; as cavins al., for caveolins only et detected and complexes essential Henley CAV1-containing crosslinked 2011; actually of al., morphology, are et More caveola (Hansen none formation influence endocytosis, caveola that and a interactions to dynamics contribute the the of are proteins these and region; network and although EHD2, However, neck caveolae, Syndapin2). known ATPase as (alternatively narrow with PACSIN2 -like protein the associated the BAR-domain-containing at GTPase, be particular, dynamin to in have found properties concentrated, been -remodelling and also 1B). (Fig. membrane-binding of 2013) inherent al., coats et al., Ludwig striated et 2014; (Gambin al., the proteins et cavin Kovtun suggest polymerised 2014; fact, strongly in structure are, which caveolae cavin X-ray into assembly, and insights EM and new both using provided by have proteins crystallography cavin the of studies Recent nadto ocvoisadcvn,svrlohrpoen with proteins other several cavins, and caveolins to addition In ´ ta. 02 ej ta. 01.Mroe,rcn analyses recent Moreover, 2011). al., et Senju 2012; al., et n 1,2, n rt .Collins M. Brett and * 1, * 1269

Journal of Cell Science COMMENTARY oano ai1 hc epooe–mgtfr oldci tutr.Prin fteCvn eune r hw ihihigtetorpasi hu hydrophobi in conserved repeats indicates 1270 two colour-coding 1D; the highlighting Fig. shown in shown are elements residues. sequences repeat Cavin1 (green) to the the polar of unique blu of and Alignment a repeats Portions (E) by (blue) (hendecad/undecad) structure. Cavin1. surrounded 11-residue basic coiled-coil (z) residues Putative (orange), a zebrafish acid i (D) form in amino acidic five cavins; might phospholipid type. the four – in residue the and propose in implicated in Cavin1, we conserved conserved residues (h) – phosphorylatio strictly are identified which are identified red previously Cavin1, red and highlighted of in as Secondary-structure-ba residues residues sPESTfind), domain well (C) acid acid by as caveolae. Amino amino (predicted of 2012), 2014). conserved; al., domains dynamics e al., et highly PEST the caveolins et Kovtun modulate predicted with (Hornbeck 2009; can surface, are al., database et and caveolar alignment Phosphosite (Hansen neck the the the caveolar coat on from that the Indicated sites proteins with cavins. ubiquitylation associate membrane human peripheral to of are known alignment Cavins is sequence coat. EHD2 layer. caveolar membrane the interior of the model proteins. Simplified of (B) family cell. cavin the fibroblast and Caveolae 1. Fig. C zCavin1 hCavin1 D AB Cavin4 328 Cavin3 Cavin2 360 Cavin4 253 Cavin3 243 Cavin2 283 Cavin4 195 Cavin3 186 Cavin2 239 Cavin4 134 Cavin3 126 Cavin2 159 Cavin4 54 Cavin3 46 Cavin4 1 Cavin3 1 Cavin2 1 Cavin1 380 Cavin1 309 Cavin1 229 Cavin1 155 Cavin1 75 Cavin1 1 Cavin2 79 L RVKFSKMEKTKTENLEKTKTKENLEKTRVQQNIGNMRTKENLEKTRQKTKENLEKTKQKTRENLEKTR HEKNI L RVKFSKMEKTKTENLEKTKTKENLEKTRARDDFEKV...... HEKTL 230240250260 270 MGEDAAQAEKFQHPGSDMRQEKPSSPSPMP S..TPSPSLNLGNTEEARDNSNAEHK E...... PHEGRIPTPPLKVTFKSQVKVED...... DES....LLLDLKHSS ...... ETSRGSNSGMDSNIDLTIVDEEESVALEQAQKVRYEGSYALTSEEAERSDGDPVQPAVLQVHQTS D...... DAVLVKSDS...... KIAPFMRLRGDEERFKSSNSKEASKDRTVAEGEECAREMGVIIARSESLGPISLYSDE.....LSPEHEAARPVYP RAMVDPGPGAEEA....LLQESA...... KVPFVRGESSGSSPFKSL..TGRESHAENETKSEDLPS.SQMPNDQEEESFAGHSEASLASALVGEIAEEAAEKA KVPFVKIRGDARSTAYKF..THEQVEVLKATEMVEVGA.DD.EGGAERGEAGLRRGS.....SPVHALLEITEES AEIDNIKKFSTRNLDKKVNRITRIVTRRERLRRQ...... KENMQKQRPGRLRQSGERLSG AEVSLRRSKRLGSATQL...... GRKGPAAPPPTPVPPP.GRAQPEQPALPTLEPEPPQDEE AEVDSLKKFSRNIEKKMNKLGTKIVSVRREKIKSHQ...... QK...... LTSNKI AEVDKKFTTRTLEKRMNKLGTRLVRREKLKSDHDFSKEKMEKTKVRTRENLEKTRLKKENLEKPRKLRKSFTPDVVY PESRIFQESLVEDDVDLSSDEEVERLRKSGHKFRCTSVKDRNLTENQEDD...... IFDPP...... YYSAKE PESRLFEEVASAEGSSDEARLRRTGLKGEFQKAPEPLGPADQSL...... QLEAE...... EPVQ PESRIFQEEIASVVEEPDEDLDEVEAKIKRSLKNEFKQPVSGAVEGKNKSLEETLHTVDLSSDDPHEALEDSAES PESRIYQDELAIEGEELELSSDEEEIERIKRSGLRVKKSSKSLKESEALPKE...LGEGERPEEDAAAAVE...VV RQLTKVSEVIEMEIKDSHNINKRKAHIKVKRVEKQIVKKVIMKKKFRHRENAQSGHIFAKQEE RQLTKVSEVGVRISRHSNTLHAARAAQVRLANLVARGGSSGGADAQAENAQEHGL RQLTKVSEVMELEGVKIDSKSSNTVSKLERKAHRMDRQAQVKRLNLLRRHFKISGYQAACHAQ RQLTKVSEVLEMEGIHSNTVSKRKVKVRLERQQIKKLVNLLRRFKMAEAQSGS LKQNNAALTIVTVDVASIVDSVQS...... MEHGSASDKIHQNRLSSVTEDEDDAAKR LKQSAAVAVTVVTLELASMLETL...... MRELER...GPVPEPAGGPHRERGG LKQSIQVAVTVLTLDLVNMLDVQN LKQSQAQVVVLSLDIIAVDQIQT...... MEDPTLYIVERPLPGYPDAEAPEPSAGAAEEPSGAGSEEIKSDNGGAQ Key HR1 eet1 eet2 eet3 eet4Rpa Partial 5 Repeat Repeat4 Repeat3 Repeat2 Repeat 1 Phosphorylation site Predicted PEST domain A lcrnmcorp fcvoa ub secvoa akdb e otdln)a h elsraeo a of surface cell the at line) dotted red by marked caveolae (see bulbs caveolar of micrograph Electron (A) Residue implicatedinphospholipidbinding Ubiquitylation site HR2 HR1 PtdIns(4,5)

NTGSAGEAERN ora fCl cec 21)18 2917 doi:10.1242/jcs.167866 1269–1278 128, (2015) Science Cell of Journal P 2 -binding repeat PM Key HR1 E 238-267 hCavin1 252-315 zCavin1 HR2 Caveolin Cavin EHD2 LEKTR LEKTKQKTREN LEKTRQKTKEN LEKTRMRTKEN LEKTKQKTKEN MEKTKQKTKEN SKGN LEKTR LEKTRLKTKEN MEKTKVRTREN KEK nteractions bde in mbedded (yellow), c o are box e h HR2 the and n man sed

Journal of Cell Science aiyhsoiial endsrbdi tde mlctn them the implicating of studies in member described each been caveolae, originally are has cavins of family mammalian components four all recognised that now fact the proteins despite coat Remarkably, of family cavin family. The into protein caveola insights cavin molecular of the emerging of model the properties refined unique on a the based is propose that and formation data structural recent COMMENTARY aishv ensont neg xesv post-translational extensive are undergo to that shown been 2008). organisms have al., et cavins these (Kirkham caveolae in of formation roles the to suggesting unrelated and worms to insects, in can specifically observable likely caveolins species, some non-vertebrate contrast, morphologically in is In found be 2010). of and also Nichols, appearance and vertebrates, predicted (Hansen the to caveolae of and limited with conservation profile is coincide genomic sequence orthologues The acidic cavin structure. highly secondary a level, sequence are DR1, disordered share the as HR2 at (Gustincich conserved to do and well refer HR1 not but HR2 we are 1C). these (DR) (Fig. DR3; and regions and 2014) DR2 disordered 1 delineated al., three et by (HR) clearly Kovtun separated levels. region 1999; two al., structural helical et within conserved and called and elements regions sequence homology structure at sequence secondary share family cavins constitute protein cavins All The unique 1). Box (see a processes cellular unrelated in ytmwsaotdfrtepoen rmCvn oCavin4 to Cavin1 from 2009). al., proteins et McMahon the naming 2009; common al., for et a (Bastiani adopted that suggested was was and system it with and associate common that caveolae, and proteins stabilise similarity membrane sequence single peripheral of a as into grounds function united the therefore, on were, family proteins McMahon protein four 2009; associate The al., Cavin2/SDPR, 2009). to et al., Hansen and 2009; found et 2008), al., also et Pilch, (Bastiani were caveolae and with Cavin4/MURC Liu al., 2008; was and et these al., (Hill Cavin3/SRBC Cavin1/PTRF function of et and Subsequently Liu formation grouping caveola 2008; family. suggested for required 1998), single be to al., shown a (Izumi et observations into earlier Mineo proteins with 1997; together the al., taken (Aboulaich et given that, SRBC 2004) proteomics-based was and al., with protein PTRF et coincided caveola-associated the of data caveolae, identification These later of ‘Cavin1’. which constituent to found 2001), moniker was main PTRF al., As a in 2005). et al., abundant be et (Vinten (Vinten is PTRF types that as identified cell protein was 60-kDa several a of of caveolae discovery the with began 2008). coiled-coil al., muscle-related C et name Ogata kinase official (MURC, protein protein the in as received C- identified known and initially to muscle, was officially binds Cavin4 now (PRKCDBP). that protein is delta-binding product but gene (SRBC) SDPR-related kinase as named function and its and 1997) for C kinase discovered protein for first (Gustincich adaptor an was name Cavin3 (SDPR) official 1993). the Schneider, given protein was protein and in a serum-deprivation-response as deprivation (PS) described serum PtdSer later initially was upon for was It induced affinity 1990). SDR) al., high et PS-p68, and (Burgener as platelets abundance (Jansa known its (PTRF) (also for factor Cavin2 discovered official release 1998). a the transcript al., as and receiving et identified I termination, first polymerase transcription was name in BBP), Cav-p60, involved family as factor protein known cavin the (also defining Cavin1 and Discovering 1. Box hra hi ucinlsgiiac ssilntcer all clear, not still is significance functional their Whereas h dniiaino aisa tutrlcmoet fcaveolae of components structural as cavins of identification The d oooost DR(zm tal., et (Izumi SDPR to homologous a -helical ellrsgaln ahas n h oiiaino aeleis caveolae to of coupled mobilisation intimately the Kru and is 2013; pathways, regulation adipocytes signalling al., their cellular et that of Humphrey suggesting 2011; 2008), stimulation al., et following be and sites to (Aboulaich found Hansen phosphorylated been identified have in Cavins experimentally 2014). rapidly (reviewed al., et DRs of Kovtun 2010; the dozens Nichols, within turnover accelerated with primarily cavin cavins, distributed modification of in abundant all an ubiquitylation is Cavin2 observation of Phosphorylation for 2012). al., role non-caveolar the et (Breen potential in of a and involved suggests degradation 2014), are (Williams proteasome-dependent cavins Palmer, DRs and in identified SUMOylation flanking been and 2014). also microtubules have and al., sites with et ubiquitylation 2008) (Kovtun association oligomerisation Pilch, to cavin C- and leads the of region (Liu removal cavin proteolysis DR3 because limited cavin terminal This of of 2004). localisation al., modulate regulator might et (Aboulaich regions important of PEST amount significant cleaved an a is Cavin1 Interestingly, of is function. sites and and homeostasis that degradation suggests family sites this these of protein of abundance feature in DRs The characteristic sensitivity. the proteolytic a enriched in residing are (sequences threonine) cavins and of motifs serine acid, PEST glutamic proline, 1C). (Fig. modification Aolihe l,21;Aolihe l,20) oee,the question. However, unanswered phosphorylation important 2006). extensive an this al., remains membrane of et plasma significance Aboulaich the functional 2011; from precise loss al., its (Bai et with (Aboulaich association associated phosphorylation is Ser/Thr caveolar at Cavin1 insulin-induced of DR3 enhance Likewise, 2011). within can al., Cavin1 et Ser366 of phosphorylation and/or one phosphorylation for these Ser365 except suggests assembly, of caveolae that to None study linked 2002; been 2005). al., yet have et Zerial, events (Pelkmans and activity kinase Pelkmans to connected reportedly motneo nesadn h ifrn oe fcavin of modes different the understanding tuning oligomerisation. in the of isoforms highlights 2013). and cavin al., functions importance of et tissue-specific for importance (Hansen properties low- the levels heart, caveola not low suggest of but at data lung accumulation expressed These in is caveolae the knockout endothelial Cavin2 Cavin2 of of where addition, loss result the In in in of oligomers. the a resulted seen size cavin average as are for mass smaller Cavin2 complex molecular a required with of cavin correlate and levels they cavin’ the tissue; High ‘gatekeeper fat expressed caveolae. and a lung ubiquitously all considered of is be formation can it Cavin1 Hansen because 2013). 2009; strikingly al., al., et cavin demonstrate (Bastiani the et types patterns of expression cell abundance the cavin relative and different the between tissues by ratios Various tuned is isoforms. expression stability coat and cavin the oligomerisation the the of distribution to that weight suggesting sensitive the cavins, 2010; different of be al., variability et to This (Hayer appears 2013). 60S al., is and et sedimentation 40S pool Ludwig apparent between cavin with In varying 2013). complexes main coefficients al., large et the into Ludwig lysates, 2014; incorporated al., cellular et and 2013; Kovtun detergent-solubilised al., 2010; homo- et al., Hansen order 2014; et of al., Hayer et higher independent (Gambin caveolins into are or membrane interactions associate cavins Cavin–cavin to that is hetero-oligomers. ability observations consistent the strikingly possess most the hetero-oligomerisation of and One homo- – subcomplexes Cavin ora fCl cec 21)18 2917 doi:10.1242/jcs.167866 1269–1278 128, (2015) Science Cell of Journal nsitu in Vne ta. 05,peual tthe at presumably 2005), al., et (Vinten gre al., et ¨ger 1271

Journal of Cell Science ais(abne l,21) hc vrl orltswl with 2005). Zerial, well and Pelkmans correlates 2013; overall al., et which (Ludwig 2014), reports previous al., 50 approximately et caveolar and (Gambin the subunits cavins that caveolin 150–200 suggested comprises CAV1 coat with to of oligomers Cavin1 analysis cavin The coincidence various single-molecule 2013). addition, al., In et be cavins. other to Ludwig estimated 2014; was al., ratio CAV1 et (Gambin 1 to on oascaewt ihrCvn rCvn tartoof ratio a at was Cavin3 Cavin1 or Cavin2 complexes, either isolated with associate In to coat. found caveolar the of members. functional analysis family unique structural other confer to to this able relative possess is Cavin1 it on indeed, whether whether properties determine so, does, to if domain and, interest element of HR2 be Cavin1 therefore, 2000) the will, al., right-handed et It Stetefeld a 1E). 2001; into (Fig. al., fold et i.e. (Burkhard to undecad), structure suggests found highly coiled-coil called repeats previously (also five repeat, the hendecad with 11-residue, a of comprise insertion, Alignment potentially longer they repeats. is Zebrafish even tandem 1D). Cavin1 (Fig. an conserved human eleven zebrafish B.M.C., possesses when from of and orthologue Cavin1 clearer elements its even (O.K. to tandem-repeat compared becomes 1C) the two This (Fig. that of residues. reveals domain consists examination HR2 sequence Closer the in observation). the sequence unpublished inserted not of an Cavin1 of of for presence feature centre caveolae unique the required form One is member to cavins. noticed able other previously family being of as absence only well the the as in caveolae, is property exclusive all particular of it this It formation mutually of because 2013). basis al., Cavin1 underlying into et the of 2014; Ludwig is segregate 2014; what al., unclear al., is and et et (Gambin Kovtun for domains Cavin1, compete caveolar 2014; Cavin3 to and al., Cavin2 Moreover, binding et 2013). associate be al., (Gambin to et unable Ludwig other to are 2009). highly Cavin3 each the and order al., with with revealed Cavin2 hetero-association, that et in co-expression and exception major homo- (Bastiani following of Cavin1 patterns coat cavins promiscuous require caveolar of cavins the al., Analysis et into other (Hayashi 2008; patients incorporated al., All Cavin1-deficient et (Hill and tissues 2008), 2009). all al., in et caveolae which of Liu animals, This loss 2008). Cavin1-knockout complete al., of a et studies show Hill with 2013; formation agreement al., with caveolar in et trigger engage is Hansen 2009; to to al., membrane et ability plasma (Bastiani Despite the the 2013). at have cavins, al., caveolin homo-oligomers reported different which et Cavin1 of complex Ludwig 2014), properties only 2010; homo-oligomerisation cavin al., similar al., the 60S-size et et (Hayer (Gambin the previously monomers on represents containing cavin complexes potentially protein 50 of formation average the correlation in fluorescence results that oligomerisation single-molecule co-translational, probably using rapid, revealed approaches models cell or COMMENTARY vrl eodr tutr rdcin niaeta cavins 1272 that indicate 1997). predictions al., predominantly et structure Izumi are 1999; al., secondary et 2004; Gustincich Overall al., 2009; et al., (Aboulaich et structure Bastiani supercoil a Early of zipper unique structure. leucine suggestive and N-terminal apparently motif, potential architecture a an their identified analyses of comprise sequence terms cavins in family the protein above, outlined the of As structure – domain era oligomerisation molecular cavin the enters coat caveolar The eetsuispritdaqatttv rti composition protein quantitative a permitted studies Recent bevtoso aiu aisepesdin expressed cavins various of Observations a hlclpoen,wt ihcnetof content high a with proteins, -helical , o4adwsidpnetof independent was and 4 to 1 nvitro in systems , 2–3 ai1floigcosikn Ldi ta. 03 (Mohan of 2013) nature immediately al., trimeric domain et apparent HR1 (Ludwig the crosslinking cavin for following the explanation Cavin1 X-ray First, of an coat. nature cavin provides 2015). the trimeric of has assembly the and al., the structure motifs for highly This leucine-zipper implications and important 2A). predicted et (Fig. shorter conserved the coiled-coil a incorporates Mohan trimeric reveal parallel mouse Cavin4a both extended 2014; zebrafish from domain and interactions HR1 Cavin1 al., the cavin–cavin of structures functional crystallographic et underlying domain readily this (Kovtun interactions that these and heteromeric indicating proteins, drives and form full-length the homo- homogeneous for observed specific a fragment the this importantly, in Most recapitulates 2014). and isolated al., recently et (Kovtun has oligomers be crystallised it can larger but HR1, 2009), form an al., that to found et been (Hansen tendency degradation their undergo by hampered cavin all species. by in all seen together in is This and linked 1C). isoforms, conserved whereby are (Fig. DRs sequences proteins, extraordinarily charged negatively HR cavin acidic, an charged the is positively to basic, There pattern 2014). electrostatic alternating al., negligible et and (Kovtun sequence disordered/extended nipratfnto.Cvn sa xeto nta tpossesses it that in exception suggests an conservation is crucial. its Cavin2 cavin–cavin are and function. unusual important highly 2C), residues is an of (Fig. coiled-coil which structures the yet of coiled-coil specificity centre in clear the at differing not pair His–Thr is The the it but will to heptads interactions, these contribute of coiled-coil sequences also unrelated The chain interacting. excluding side from Gln other thus sequences equivalent protomers, in an conserved to adjoining several residues pairing are from require these there that cavins, of residues In Gln of below. conservation specificity indicated Cavin1 and is the the of cavins shown, defines residues core are what the 2B, HR1 often hydrogen- Fig. is In of formation. this capable coiled-coil side-chains and of pairing, inclusion bond the with interface vary the at can the residues However, central structure hydrophobic protomers. the the between a defines of which labelled by centre repeat, acids, amino heptad the characterised seven at of are repeat lying pattern chains Coiled-coils side pair 2B,C). Thr (Fig. hydrogen-bonded and conserved His a of share cavins First, observed. complexes. cavin the of architecture overall high- ( assembly folds by rod sequence elongated and cavin confirmed and each rigid of highly be fraction a the into large to Third, a assemblies. that remains HR1 shows structure mixed it Ludwig of 2014; studies although structural al., resolution 2013), et (Gambin al., Cavin2 complexes of et molecule caveolar per in the Cavin1 of Cavin3 indicating copies structural or three studies to the biochemical two of both previous presence satisfying the or be to and Cavin2 explanation appears constraints either reasonable This with arrangement. most replaced exclusive the mutually is a coiled-coil the in whereby the hetero- Cavin1, Cavin3 are of of copies helix Cavin1–Cavin3 the two of third composed and Second, be to Cavin1–Cavin2 likely below). most by formed these (discussed oligomers how question an although remains must blocks, polymers important coat membrane-associated building cavin into the trimeric assemble that trimers is of this composed of consequence be The 2015). al., et slto fcvn srcmiatpoen sgenerally is proteins recombinant as cavins of Isolation w te oal etrso h R oldci eealso were coiled-coil HR1 the of features notable other Two , mi it)ta,a icse eo,i eeatt the to relevant is below, discussed as that, width) in nm 3 ora fCl cec 21)18 2917 doi:10.1242/jcs.167866 1269–1278 128, (2015) Science Cell of Journal , 0-eiu ai rget h N-terminal the fragment, cavin 100-residue a and d oiin fti repeat this of positions abcdefg , b 6n nlength in nm 16 setcontent -sheet emdthe termed ,

Journal of Cell Science COMMENTARY fukontplg,peual rvnb self-association by secondary in driven increase This trimerisation. presumably HR1 an through topology, established adopts it unknown However, to isolation. HR1 of contrast in with in unfolded that, is combined found least) when at was length Cavin4 It helical in (of nature. vary HR2 putative in can HR1, basic sequence second highly cavin whose also The HR2, is order is but 2014). cavins higher the al., of of domain et assembly are understood (Kovtun the cavins poorly oligomers support the through can within and, mechanisms, domains important functionally Other required not clearly formation. complexes is larger caveolar of it formation for specificity, the cavin–cavin promote to determines sufficient HR1 proteins membrane cavin the the Whereas in of structures role order discussed Higher important As an polymers. residues. 2A), cavin play Arg (Fig. the by and C-terminus residues association Lys its these of at below, clusters surface of of basic has regulation composed domain conserved HR1 of cavin highly level the Second, a additional ubiquitylation turnover. and an multiple assembly suggesting cavin contains and, 1C), also cavins (Fig. sites the region to assembly. their of this it regulating structure for Remarkably, speculate mechanism stable a we will the represent potentially, residue and for this pairing, consequences of have hydrogen-bonded interactions. phosphorylation the al., Clearly, et disrupt cavin–cavin Kovtun 1C). is 2013; it (Fig. al., that et residue 2014) fact (Humphrey on the structure Thr despite the cavins, within the the buried several this determine that in phosphorylated to indicate be interesting can of studies be proteomics will Interestingly, impact it and indica dimensions His, with structural element of rod single instead micro a of electron Gln comparison. representation g stained are schematic for Negatively that a scale (D) seen shows to coil. are inset hydr shown elements The coiled the rod-like is topologies. showing the of curved coil structure of Arrays Cavin4 and Cavin1 coiled layer removal. and linear the HR1 lipid hydrophilic both Cavin3 of and display central Cavin2, section crosslinking but Cavin1, a Cross representat after dimension human (C) creating ‘worm-like’ protein in of Cavin1). in full-length chains, residues in domain Cavin1 side corresponding Thr105 HR1 expressed His–Thr Cavin1 and the bacterially central mouse (His102 this, recombinant the pairing the Below of His–Thr chains. between structure central side pairing the the as bond shows highlights shown panel box repeat top represe dashed heptad The surface The the proteins. the shown. of cavin shows the residues panel of Bottom central repeat C-terminus. the heptad to with The N- (B) (red from domain. potential red HR1 to electrostatic the blue for coloured coloured representation structure EM. ribbon same and in the crystallography coiled-coil by homotrimeric proteins the cavin shows the panel of Structure 2. Fig. AB C H102 T105 C A T105 C H102 H102 B T105 A B D a hlclconformation -helical 52 Cavin4 A..I...L..V...V..V... Cavin3 A..I...L..L...L..L... Cavin2 A..V...L..L...L..V... Cavin1 G..V...L..I...V..I... . ,blue V, 0.5 a..d...a..d...a..d...a..d...a..d...a..d...a..d...a..d...a..d...a..d...a..d...a..d...a..d...a..d A rsa tutr ftemueCvn R(otne l,21)(D D4K) Top 4QKV). ID (PDB 2014) al., et (Kovtun HR Cavin1 mouse the of structure Crystal (A) 5 05V.Nt h eino togpstv hre(le ihnteCtria afof half C-terminal the within (blue) charge positive strong of region the Note V). +0.5 ehp uhls otiuinfo h membrane-embedded with the assumed. cavins, fact, from previously in as membrane-associated contribution are, caveolins less caveolae peripheral of much the feature perhaps defining 2013). by a al., densities as formed striated et the accepted Ludwig that are certain 2014; that almost al., seems of now et surface it (Gambin the Altogether, on bulbs striations caveolar by membrane suggest the coat strongly peripheral analyses cavins adopt the that ultra-structural they on that specifically recent elements focused that Furthermore, striated EM using 1992). the cavin al., to these et observed of similar morphology as strikingly and caveolae, size is distinct The 2014). a rods al., possess et to (Kovtun revealed structure, are cavins rod-like purified extraction, the detergent and however, of crosslinking gentle structures Following diameter. spherical relatively display complexes isomorphous al., protein is et relatively homo-oligomeric (Kovtun purified lipids that The of quantity 2014). substantial assembly a with co-fractionate larger even the i.e. size, an flank in heterogeneous that of the in DR3 formation C-terminal mass and and molecular N- the a DR1 of with inclusion Interestingly, complex step, range. protein sub-MDa oligomerisation large additional a an yielding with coincides also structure flpdrf,wt hrceitcivgntdtplg,ada and topology, al., et invaginated (Ortegren species characteristic lipid various a in enriched species with leaflet membrane distinct raft, a comprise lipid to of considered generally are lipids Caveolae membrane with interactions Cavin hnioae rmbcei,rcmiatcvn r on to found are cavins recombinant bacteria, from isolated When Q Q Q Q ..I... ..L... ..M... ..L... ora fCl cec 21)18 2917 doi:10.1242/jcs.167866 1269–1278 128, (2015) Science Cell of Journal H Q Q Q ..L...V..V...L..L... ..L...V..I...L..L... ..L...V..I...L..L... ..M...V..I...L..L... , 3n ogad5n ie(i.2D) (Fig. wide nm 5 and long nm 23 nvivo in 55 A , H H Q H ...... – MDa. 1–2 T T T T ...V..L...... 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Journal of Cell Science Kvu ta. 04 Fg C ) h aewr a also has work same The PtdInsP 3). HR1 caveolar 1C, between extent, (Figs connection lesser 2014) a a the established al., to of et and, composed domain (Kovtun are HR2 polybasic that conserved regions PtdSer-binding on two revealed independent data have recent cavins but truncated 2009), systematically of of al., end affinities et lipid the McMahon at 1997; was al., region et site conserved (Izumi PtdSer-binding a HR1 2008; A in 1998). al., reside al., to et postulated et Hill previously Mineo 1999; 1997; al., al., et et (Gustincich Izumi family the of members o ntr,ehnetelclsdercmn fPde and PtdSer of enrichment localised the enhance turn, PtdIns(4,5) in likely are to, cavins be of phospholipid-binding polymerisation independent to and association two membrane thought sites, least each are that at and trimers possess 2014), will cavin al., et trimer 20 (Gambin vesicle and caveola each 15 Given in present between may 2014). that al., HR2 et fact that and (Kovtun the thought HR1 phenotype intermediate is between this it binding explain HR2, and membrane localisation, in or membrane redundancy to 2014). in al., Cavin2 HR1 an block in et result absolute or not Kovtun either does individually 2009; residues Cavin1 these al., in mutating However, of et (Hansen residues recruitment membranes caveolar the Lys the affects of and mutation substantially concomitant Arg phosphoinositide that shown of conserved been recognition has It specific headgroups. somewhat general a HR1 a possess but membranes, promotes cavins phospholipid that charged within negatively is for proposed site affinity thus, phosphoinositide-binding model, The high-affinity HR1. a revealing oriae sebyo ltrncae noyi vesicles endocytic 2014). al., et -coated Kelly 2011; al., of et the (Antonescu being example assembly prime al., the with et coordinated 2002), and (Li al., et proteins association McLaughlin membrane-associated 2014; membrane of in regulation involved allosteric often are these of phospholipids headgroups exposed Cytoplasmically membrane 2014). plasma al., the et (Ariotti within domains PtdSer of re-organisation results 1996). CAV1 Casey, in of and knockdown Pike by 2009; caveolae al., of lipid depletion et Furthermore, Fujita distinct 2011; al., physiologically et (Fairn forming pools caveolae, in concentrated 45-ipopae[PtdIns(4,5) phosphatidylinositol phosphoinositide (4,5)-bisphosphate the of and plasma the PtdSer presence addition, of membrane, phospholipids In the charged 1992). negatively al., abundant requires et most Rothberg specificthe particular, 2012; their al., et in on (Breen dependent cholesterol and, is composition caveolae lipid of assembly The 2004). COMMENTARY 1274 for essential is and 2009) al., et Yao 2004; al., membrane et caveolar (Ortegren domains of component abundant another is Cholesterol Cholesterol reports Further 1990). al., Ca et similar (Burgener cells identified platelet PtdSer-binding of major a protein as and isolated originally protein and was distinct assembly Cavin2 caveola function. the control to how cooperate caveolae to of species as lipid arises therefore, question, The Phospholipids vrl,hwvr ept h la ciiyo aisin cavins of caveolar in activity lipids these studied. poorly clear of quite role remains the function actual the 2013). despite al., binding, et phospholipid Simone however, in 1997; al., involved in localised Overall, et are important (Izumi with transport that be increased and factors then signalling cavins caveola-associated may and which other lipids, of regulating those phospholipids of association concentrations charged between negatively mechanism feedback P 2 bevda aele hssget cooperative a suggests This caveolae. at observed 2+ idpnetPde-idn ie nall in sites PtdSer-binding -independent P 2 aebe ugse obe to suggested been have ] 2 n cavins, and h aelrdmi,ptnilyrsligfo perturbed Murata 2013; from al., resulting et 1995). al., potentially et (Hulce interactions domain, caveolin–cholesterol caveolar within bilayer lipid the the of general organisation more physical a the disassembly reflect of perturbation to cavin likely and are flattening depletion cholesterol caveola al., following et that (Epand suggest proteins We cholesterol-interacting 2010). fide – cholesterol bona with in associate to seen membrane into as lipid insertion the of for layer required acyl are the that stretches hydrophobic obvious cholesterol-containing explanantion to significantly the membranes bind be not do direct to cavins A unlikely because clear. not is required is interaction localisation is in cavin–cholesterol cavin cholesterol the and why (reviewed formation in reason caveola cholesterol the caveolae for 2005), cellular implicating Parton, literature of and Martin of Although homeostasis 1992). history the and al., rich into transport et a cavin Rothberg is of 2012; there dissociation al., et and 1998; (Breen flattening cholesterol al., cytoplasm of caveola et depletion in Hailstones Pharmacological results 1992). 2012; al., al., et et Rothberg (Breen formation caveola ael eeain n umutn hsbrirrqie the to requires barrier coat. barrier a cavin-caveolin the this present of surmounting assembly cells, coordinated and mammalian generation, in caveola the of membrane key properties that biophysical plasma suggests and/or the This lipids concert. proteins, structure, in either factors, act membrane proteins cavin shape and caveolae caveolin to morphological cavins of of formation and abilities individual caveolins the both despite peripheral However, domain-containing proteins. ENTH molecules, membrane and curvature-inducing BAR-domain- membrane as of such family broader the vitro aeln aea neetaiiyt eeaeshrclparticles spherical generate to ability inherent an have membrane-embedded caveolins bacteria, in remodelling expressed membrane heterologously in When cavins of role The ebaermdligadcraue n oyei scaffolding polymeric and membrane curvature, promote and with proteins remodelling specialised interact membrane ‘cargos’, transmembrane adaptors and and nucleation, lipids McMahon and 2013; promote al., recruitment small including et proteins general, (Faini In 2004). assembly Mills, of organisation of structural mechanism terms requirements, and membrane in and characterised protein well their relatively clathrin and been COPII assembly COPI, have the caveolar machineries, as in such coat, cavins vesicular of of types role Many the for model A and cavins purified when and tubular 2009), extended al., mammalian PtdSer–PtdIns(4,5) generate et in synthetic overexpressed (Hansen to when cells both membrane membranes structures, caveolar membrane remodelling biological of are invagination complex of Cavin1, the particular more in stabilise domains of and, to cavins increased bending required the to lead Instead, upon that membranes. principles cost the of provides energetic 2013) description al., review excellent et recent an (Stachowiak A colleagues of bending. and an membrane Stachowiak because and by of themselves membrane probably cost the by energetic of cells, properties increased cannot biophysical mammalian the in caveolins in differences Nevertheless, caveolae (Verma 2010). generate Cavin1 by al., in suppressed be formation can et tubule activity this induces and overexpression models cell CAV1 (Walser 2012). caveolae mammalian al., to similar et is that morphology a with Kvu ta. 04.Ti lcstecvn imywithin firmly cavins the places This 2014). al., et (Kovtun ora fCl cec 21)18 2917 doi:10.1242/jcs.167866 1269–1278 128, (2015) Science Cell of Journal nvivo in nvitro in nfc,lk aeln,cvn ln r capable are alone cavins caveolins, like fact, In . Kvu ta. 04 n ontcontain not do and 2014) al., et (Kovtun P 2 cnann iooe r used are liposomes -containing nvivo in eursta both that requires in

Journal of Cell Science COMMENTARY aispeetdaoet rps etbemdlfrterrl in role 3). their of (Fig. for the model formation features testable caveolae combine a structural propose and to and above summarise presented temporal cavins to spatial, attempt biochemical, we known section, mechanisms. final membrane understood this poorly coordinate still In and to different appears quite formation adaptors involve caveola to of the process the contrast, to In invagination. coupled are proteins i membrane plasma the with associated stably PtdIns(4,5) become cholesterol, only with but cluster membranes to believed charged are negatively Caveolins bind caveolins. formation. can of caveolar These presence oligomers. in the cavin function cytoplasmic cavin into for assembly model A 3. Fig. nnw.()A xaddve ftemmrn ufc,soigaseuaiemdlfrcvncvnitrcin hog naryo electrostatic of array an through protomers. interactions adjoining cavin–cavin in back for charge cavins model opposite for of speculative caveolar of release a for the domains consequences showing and and positive surface, caveolae lipids or membrane of membrane negative the flattening both the to of involving but of lead view interactions modifications, formation can expanded and posttranslational depletion, An curvature by cholesterol membrane (B) modified nucleate or unknown. extensively then stretching are can membrane Cavins This as particular. pool. in such cytoplasmic lipids stimuli, charged Certain negatively bud. the of caveola concentration local the increase further HR1 trimerisation A Cavin3 Cavin1 Cavin1 Cavin1 Cavin1 Cavin1 Cavin2 Cavin1 Cavin1 Key PtdSer Caveolin Cholesterol PtdIns(4,5) heterotrimer Cavin1–Cavin1–Cavin3 heterotrimer Cavin1–Cavin1–Cavin2 homotrimer Cavin1-Cavin1–Cavin1 P 2 Phosphorylation? Ubiquitylation? Cavin trimersformed Stimulus- and/orstress-inducedflattening No caveolin Cavin oligomersreleased B R DR3 DR1 – A h ai iecceivle nta rmrsto hog h R oanadsubsequent and domain HR1 the through trimerisation initial involves cycle life cavin The (A) PtdIns(4,5) PtdSer R R HR2 DR2 HR1 +– P Assembled caveolae 2 PtdSer + Cytoplasmic cavinoligomers Electrostatic protein–membranearray ai otcmlxi eivdt sebei h cytoplasm peripheral the The in 2005). assemble to al., believed et is complex (Pol coat lipids, al., cholesterol on cavin relies et also particular (Hayer plasmalemma cavins the in to with export interaction Caveolin and 2010). independently formation membrane caveola plasma caveolin the of in to results directed and cell, being the oligomers of compartments exocytic early in – Caveolin oligomersclusteringwithPtdIns(4,5) aelnoioeiainadisrini h ebaeoccurs membrane the in insertion and oligomerisation Caveolin P 2 n tSr(hw ttetp;sbeun erimn fcvn will cavins of recruitment subsequent top); the at (shown PtdSer and ora fCl cec 21)18 2917 doi:10.1242/jcs.167866 1269–1278 128, (2015) Science Cell of Journal aelnoioe Caveolin oligomer Caveolin oligomer Non-caveolar functions? Membrane curvature Polymerisation andlipidclustering P 2 , PtdSerandcholesterol Cavin recruited ainaestill are mation h distinctive the nothe into 1275 n

Journal of Cell Science oto rcse uha rncitoa regulation, transcriptional of as upstream levels of such expression importance – relative the by of highlighting processes defined degree thus control coat be high caveolar cavin, cavin the to functional a in each of ratios likely suggests cavin assembly consequence, are the this a in As Overall, complexes. involved with cavin is cavin DR3. of coincides cooperativity regions step and Last, C-terminal this final and N- DR1 the 2013). and disordered whereby the domain requires al., cavins, oligomersaion the HR2 of et of the oligomerisation formation Ludwig within the 2014; their 2014; structures al., induces al., et dictating (Gambin trimerisation separate et caveolae thereby of morphologically Kovtun surface the and prohibited, on biochemically subcomplexes strictly Cavin2–Cavin3 into of segregation is formation the either complexes Third, Cavin1–Cavin1– of or copy respectively). one (Cavin1–Cavin1–Cavin2 Cavin3, and with Cavin3 Cavin1 and or of Cavin1 association, Cavin2 copies and two Cavin2, cavin–cavin a containing and is Cavin3 Cavin1 the there of Second, consisting of N- 2014). heterotrimers the al., stoichiometry in et structures (Kovtun specific coiled-coil domain their HR1 now of terminal heterotrimers are and association principles homo- the key form through proteins some yet cavin above, in the not outlined exact First, is present emerging. as The recruitment but, 2013). molecules and clear al., oligomerisation cavin fully et cavin Ludwig of of 2014; number mechanism al., et the (Gambin of to caveolae formation similar the in 2014), average on results contain that and multimers autonomous is oligomerisation hyaems ieyt xs na qiiru ewe a between PtdIns(4,5) equilibrium as and an such the (PtdSer) in phospholipids, on phosphatidylserine plasma-membrane-enriched exist dependent of state, to presence membrane-associated likely and most cytoplasmic are abundance regulate they caveola to over – control exert degradation function. and ultimately, protein and, and cavins transport processing, aeln ensgetdt eiprati idn to binding in in PtdIns(4,5) organisation important and their PtdSer be regulating cholesterol, to and including microdomains, lipids suggested membrane been the various in long factor Caveolae formation. important have caveola most for the requirement cavin–caveolin is dual interactions lipid and protein driver caveolae. primary to the recruitment as cavin interaction of argues purified protein–protein which direct between data), a unpublished association against been (our proteins have stoichiometric caveolin we a and although cavin a 2015), detect some al., play to et may is (Mohan unable that process There this association in buds. cavin–caveolin role direct the shaped for form must spherically evidence to cavins invaginated, caveolins biogenesis, membrane-embedded typical most caveolar the the with of perhaps remains cooperate aspect what al., in understood et to but, Kovtun 2012) poorly abilities 2009; al., et intrinsic al., Walser et possess 2014; (Hansen caveolins structure membrane and reshape cavins Both 2014). ro oascainwt aelno h lsamembrane, plasma the on caveolin observations with and association to prior COMMENTARY 1276 or is cholesterol caveolae either for of lipids depletion these 1996; that Casey, of sense PtdIns(4,5) and affinity the Pike The in 2013; 2014). reciprocated Pozo, Murata al., 2009; del al., et and et Yang Parton Fujita 2011; 1995; al., al., et et Fairn 2014; al., et (Ariotti fcvoa rtis(re ta. 02 ohege l,1992; al., et Rothberg 2012; al., et (Breen proteins caveolar of ae nbohmclaaye fioae eobnn cavins, recombinant isolated of analyses biochemical on Based epooeta h omto facoeaientokof network cooperative a of formation the that propose We P 2 a,i un edt oso aeleo dissociation or caveolae of loss to lead turn, in can, nvitro in n nclssgetta cavin that suggest cells in and , 0mnmr Gmi tal., et (Gambin monomers 50 a P hlclsecondary -helical 2 Kvu tal., et (Kovtun P 2 n tblsstecaatrsi opooyo aele The is caveolae. caveolae form of to caveolins to morphology binding cholesterol characteristic membrane, of membrane importance the the the al., bends at stabilises caveolin and complex et and cavin of cavin Phospholipid action (Ariotti the cooperative the 1997). signalling accommodate to To al., reported Ras 2014). of also et lipid-raft-associated was localisation formation modulate (Izumi caveola 2013), for by al., caused activities et required redistribution (Simone enzymatic EHD2 be as or such also components, cavin protein may the concentration other of elevated phospholipids resulting trapping’ The ‘electrostatic of membrane. of We the type HR2). on a and complex be HR1 to (i.e. this cavins imagine in stretches membrane-binding 02 i n ic,20;Mrt ta. 95,signal 1995), al., et Murata 2008; Pilch, and al., the et (Breen Liu depletion proteins cholesterol understanding as cavin such 2012; of – caveolae, stimuli caveolae, turnover to and response of in redistribution of disassembly subsequent dynamic the assembly the and control the that the in roles mechanisms Furthermore, the in to unknown. yet role stimuli, essentially post-translation addition to are important response modifications an extensive their these and play of caveolae undergo to of cycle likely cavins life are they As modification, regulated. depletion is cholesterol as following such 1992). – al., observed et structures (Rothberg arranged caveolae loosely coats the more invaginated cavin flattened as the spiral by well around the structures as that formed packed caveolae, fact exists densely the are indeed, both by adopt flexibility, supported can that partly a least such arrays at That is organised highly coats. the clathrin with tightly polymeric contrast in and be would rigid This around form membrane. to curved coats an flexible the quite relatively such be allow may and of subunits locally between dynamic consequence would contacts whereas polymers probable elastic, cavin highly neighbouring A be membrane-associated of that 3B. HRs is Fig. lattice arrangement recognise assembled in electrostatically to illustrated an able acidic as creating the be essence in that turn, charged basic cavins, envisage in negatively we may, contact Whereas below), DRs can highly (see HR2 profile. their bilayers and phospholipid and HR1 electrostatic flexibility within surfaces cavin alternating proteins; cavin overall the to conserved allowing of answer properties the DRs general of two part their in least lies remains at question it that this Although believe we polymers. speculative, spiral highly characteristic interface the membrane the create at to assemble blocks building cavin cavin– shaped the by as bending occur. and, membrane to 1995) coat appropriate caveolin lipid al., allow the important of to et properties an biophysical bilayer reflect Murata localised to the likely 2012; modulating is in role this al., believe we et above, mentioned (Breen clear also ocnrto eea odhge hni h surrounding the PtdIns(4,5) on with enriched oligomers in be to caveolin likely Thus, than are membrane 2003). plasma al., the higher et (Wanaski fold membrane several concentration n yrpoi mn cd,psessa nrni ability PtdIns(4,5) intrinsic and PtdSer an cholesterol, possesses increase acids, locally amino to basic of hydrophobic that sequence 20-residue suggest and a experiments domain’, ‘scaffolding Liposome-based caveolin 2013). the al., et Simone ilte ntaeapstv edaklo asn additional causing increasing further loop interaction therefore, feedback raft, the PtdIns(4,5) This locally positive with caveolin enrichment a of proteins. association this initiate then cavin that will hypothesise membrane-sensing we attracts and PtdSer, and ayqetosrmi eadn o ael formation caveola how regarding remain questions Many rod- the how understand to is now question intriguing An ora fCl cec 21)18 2917 doi:10.1242/jcs.167866 1269–1278 128, (2015) Science Cell of Journal P 2 n tSrlclytruhbnigo aelnto caveolin of binding through locally PtdSer and P 2 oa to P 2

Journal of Cell Science noec,C . ge,F,Dnsr .adShi,S L. S. Schmid, and G. Danuser, F., Aguet, N., C. Antonescu, soitdwt yokltlsrcue Rctre l,2008); morphologically al., et are (Richter caveolae structures cytoskeletal Verma Furthermore, with 2002; associated al., 2013). et in Mundy Pozo, Wickstro changes 2013; 2010; and that al., al., transport et relatively known et caveolar remains alter is (Hernandez can – it stability assembly stress 2011) comment, actin al., membrane and final microtubule et a and Sinha As 2006), 2014; unexplored. al., al., et et (Gambin (Aboulaich transduction COMMENTARY rot,N,Ferna N., Ariotti, E. Maratos-Flier, and S. J. Flier, M., J. Asara, C., P. Chui, N., Aboulaich, Stra and V. A. Vener, U., Ortegren, N., Aboulaich, Stra P., J. Vainonen, N., Aboulaich, References Fellowship Development Career NHMRC Fellowship an Research [APP1061574]. by Principal BMC Senior and NHMRC [APP1058565] an CE140100036]. by in number supported Excellence [project is of Technology R.G.P. Centre and [grant ARC Science B.M.C. the Bio-Nano to by Convergent (ARC) partly Council and Research APP569542 DP120101298] Australian numbers number Medical the [grant and R.G.P. and Health to APP1037320] National (NHMRC) and the Australia from of grants Council by Research supported was work This Funding interests. financial or competing no declare authors The interests Competing 2015. al., et Mohan in included now publication, to prior interactions cavin–caveolin (Umea Lundmark Richard thank fundamental We these Acknowledgements to answers biology. provide caveolae under of currently also questions techniques EM may improving development, rapidly approaches, the imaging with instance and structures for components these caveolar the combining obvious of an that reconstitution are suggest complexes obtaining their we and Although priority, cavins of domains. structures together membrane crystal come new invaginated distinctive cavins and the generate caveolins how form to course, of to and, interface cavins coats membrane how spiral understanding the cavins, of such at full-length role remain, self-assemble of the still architecture issues on many the light Yet, as some underlying The organisation. shed caveolar understood. the to in begun poorly cavins defined, has very here well is highlighted vesicles work now caveolar are of ultrastructure constituents of primary most of structures the the where vesicles, contrast inherently COP-coated stark or In clathrin- the formation. to their their govern and that study despite principles to caveolae difficult molecular it the yet, made of have proteins And core nature the of ingredients. structures relatively flexible dynamic protein a the by core importance, defined of are set and restricted morphology a possess characteristic processes, cellular highly many of regulators crucial are Caveolae biogenesis. remarks with caveolar Concluding interactions in role whether direct addressed a have be proteins to cytoskeletal remains it however, aelerglt h aocl raiaino h lsammrn to membrane G. plasma R. the Parton, and of F. organization signaling. J. Ras Hancock, nanoscale control R., remotely the M. Wenk, regulate B., Caveolae L. Tanner, L., K. initiation, size. pit and stabilization, clathrin-coated regulates Phosphatidylinositol-(4,5)-bisphosphate release transcript mechanism. and PTRF.phosphorylation-dependent I with lipase Polymerase hormone-sensitive of Commun. Res. translocation Biophys. regulated Biochem. insulin in caveolae and of of surface fragmentation the at specific (PTRF) factor adipocytes. and release human transcript phosphorylation and I targeting, polymerase reveal proteomics ne-oo .A,Zo,Y,Hl,M . oky .L,Inder, L., T. Rodkey, M., M. Hill, Y., Zhou, A., M. ´ndez-Rojo, ice.J. Biochem. o.Bo.Cell Biol. Mol. me l,21)(eiwdi atnaddel and Parton in (reviewed 2010) al., et ¨m .Cl Biol. Cell J. nvriy wdn o rvdn aaon data providing for Sweden) University, ˚ 383 los .adVnr .V. A. 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