Vinculin Regulation of F-Actin Bundle Formation

Home , Talin (protein), Vinculin

www.landesbioscience.com 1 Tolbert, E. Caitlin What doesitmeanforthecell? Vinculin regulation of F-actin bundle formation C and Biophysics; University of North Carolina School of Medicine; The University of North Carolina at Chapel Hill; Chapel Hill, NCUSA Hill, Chapel Hill; Chapel at Carolina ofNorth University The ofMedicine; School Carolina ofNorth University Biophysics; and Email: [email protected] *Correspondence to: Sharon L. Campbell; http://dx.doi.org/10.4161/cam.23184 12/08/12 Accepted: 12/07/12 Revised: 09/14/12 Submitted: scaffold adhesion, focal bundling, F-actin Keywords: Chapel Hill, NC USA; NCUSA; Hill, Chapel Department of Cell Biology and Physiology; University of North Carolina School of Medicine; The University of North Carolina at Chapel Hill; Hill; Chapel at Carolina ofNorth University The ofMedicine; School Carolina ofNorth University Physiology; and Biology ofCell Department al. al. M Berginski VS, Swaminathan org/10.1074/jbc.M111.244293 286:45103-15; PMID:22052910; http://dx.doi. 2011; Chem JBiol properties. mechanical cell F-actin bundle formation, focal adhesion, and Commentary to: Shen K, K, to: Shen Commentary ell Adhesion &Migration 7:2,219–225;March/April 2013;©2013LandesBioscience T h e vinculin C-terminal hairpin mediates mediates hairpin C-terminal e vinculin Co mm ent vinculin, dimerization, dimerization, vinculin, a ry 1 2 Keith Burridge Keith Lineberger Cancer Center; The University of North Carolina at Chapel Hill; Chapel Hill, NC USA; NCUSA; Hill, Chapel Hill; Chapel at Carolina ofNorth University The Center; Cancer Lineberger T o

lbert C lbert E , B urridge K, et et K, urridge E , G uilluy C, C, uilluy

1,2 and Sharon L. Campbell L. Sharon and folding protein that links transmembrane transmembrane links protein that folding scaf aubiquitously expressed is Vinculin variants. these actin bundling deficientvinculin investigate further and findings ous previ on our expand will we mentary, com this In function. bundling its from distinct hairpin, C-terminal for the These results suggest additional role(s) phenotypes. cellular show different that bundling deficient variants ofvinculin identified have we Intriguingly, tion. transduc force and properties adhesion both impacts region of this deletion as cells in hairpin C-terminal of the tance impor the demonstrated We also have F-actin. for bundling and Vt dimer of the formation the for both critical culin carboxyl (C)-terminal hairpin is vin the that shown have and Vt dimer actin-induced for the evidence tional addi provided Wements. recently have fila actin crosslink to able is that dimer of aVt promote to formation believed Vt in change aconformational induces dle actin filaments. Binding to actin bun and bind both to ability the has (Vt) of vinculin domain tail The tion. transduc force arolein play to shown been more recently has and survival, cell and motility shape, cell controlling in actin cytoskeleton. Vinculin is involved the to proteins transmembrane couples it where junctions, adherens and sions V protein, found at both focal adhe focal both at found protein, adhesion cell essential an is inculin C ell Adhesion &Migration Introduction 2,3, *

------interactors. other recruit able to bind and is vinculin and released are interactions head-tail partners, of multiple binding action die by day 10 by day die embryos where studies knockout mouse in demonstrated been has of vinculin tance sion strength and motility. and sion strength adhe formation, adhesion cell regulate tions. junc adherens and (FAs) adhesions focal at cytoskeleton actin the with receptors nects the head and tail domains. tail and head the nects con region linker protein-rich flexible A domain. tail helical smaller and head helical large its between tiple interactions mul through state autoinhibited tive, and IpaA; and to α binds (Vh) domain head the partners: binding multiple able to recognize is domain notransducer by mediating the linkage linkage the by mediating notransducer amecha as it to serve enabling vation, acti promotes vinculin F-actin and talin of binding Coordinate F-actin. and talin include partners binding these at FAs, to apoptosis and anoikis. and to apoptosis resistance and stiffness decreased naling, sig paxillin FAK and increased motility, increased spreading, decreased adhesion, decreased including: of defects a number [PtdIns(4,5)P (4,5)-bisphosphate phosphatidylinositol and (F-actin) raver1,actin filamentous PKC paxillin, (Vt) recognizes domain protein and Arp2/3 complex; Arp2/3 protein and α vinexin nArgBP2, CAP/ponsin, with / β Vinculin is maintained in its inac , vasodilator-stimulated phospho , vasodilator-stimulated 1 Vinculin has also been shown to shown been also has Vinculin 12-16 27 3 / Department of Biochemistry ofBiochemistry Department 3 When vinculin is localized localized is vinculin When β and isolated fibroblasts have fibroblasts isolated and the linker region interacts interacts region linker the 2 -catenin, -catenin, ]. 22-26 Through the inter the Through 4-10 α -actinin, talin 2 Co The impor The 17-21 mm the tail tail the 11 ent Each Each a 219 ry α ------,

©2013 Landes Bioscience. Do not distribute. 220 skeleton. cyto actin the and integrins between F-actin. and vinculin between interaction of the importance the investigate further will we commentary, this In survival. and structure cell motility, cell including processes cellular to playarolemany in believed is vinculin and F-actin between C-terminal hairpin (red) and and (red) hairpin C-terminal that enables Vt self-association and F-actin bundling. F-actin and self-association Vt enables that (orange) site dimerization acryptic exposes that Vt in change aconformational causes binding talin (green) to Vh (purple) and F-actin (gray) to Vt (blue), leads to vinculin activation. tovinculin leads (blue), toVt (gray) F-actin and (purple) toVh (green) talin function of the vinculin/F-actin interaction. vinculin/F-actin the of function the study to tools specific tomore lead should interaction critical this for model alternative or refined a findings, our given Hence, formation. bundle for isnecessary hairpin C-terminal the that indicating arisen has that data contrasting and F-actin, and Vt between clashes conformational multiple micrograph, the of resolution the given validation further require may model Janssen the Janssen et al., et Janssen Figure 1.

28 Model of vinculin activation, F-actin binding and bundling. ( and bundling. binding F-actin of activation, vinculin Model Additionally, the interaction interaction the Additionally, 32 release of autoinhibitory interactions within full length vinculin via binding of of binding via vinculin length full within interactions autoinhibitory of release N - terminal strap (yellow) point into the F-actin interface. F-actin the into point (yellow) strap terminal - length protein. length full inactive the in Vh with interactions due to autoinhibitory masked partially is site that binding F-actin an Vt contains The F-Actin/Vinculin Interaction 30,32 Self-Association andF-Actin ( B ) According to the Janssen model, the the model, Janssen tothe ) According C Results inVinculin ell Adhesion &Migration 29 Bundles Upon binding F-actin, Vt F-actin, Upon binding ) As modified from from modified A) As 32 32 F-actin F-actin However, However,

dling efficiency of Vt. efficiency dling bun the enhanced C-terminus oftion the dele while bundling actin impaired strap (N)-terminal amino of the deletion that integrity and stability of Vt, stability and integrity structural the affect C-terminus the from acids amino five than larger deletions dling. bun actin do not impair deletions strap previously, shown N-terminal has group band is observed for Vt Δ observed is band dimer actin-induced of the intensity band 2 Figure in shown As of F-actin. absence and presence the on Vt Δ crosslinking we performed model).plex (Janssen com Vt/F-actin for the model a structural proposed and approaches mutagenesis and docking microscopy,tron computational elec combined al. Moreover, et Janssen actin filaments to be crosslinked. be to filaments actin enables site that dimerization a cryptic through to self-associate shown been has our actin bundling deficient deficient mutants. bundling actin our Vt on and WT concentrations of actin atarange studies crosslinking conducted we F-actin, with upon association induced hairpin ( hairpin C-terminal into the leading site) 5 (lower helix 3and of helix bottom the as well as 3(upper site) 2and top of the helix sites; Vt with attwo interacts F-actin model, 1,066; Vt Δ 1,066; of Vt integrity (Vt structural do the not to alter that hairpin C-terminal to the deletions smaller By introducing variants. deletion hairpin C-terminal fromlarger data interpret to it difficult published results. published to previously contrast in bundling, F-actin for essential are residues, five last the ular, partic in and C-terminus the that shown the presence of F-actin, the Vt the Δ of F-actin, presence the Vt Vt for and WT formed Δ are cies spe trimer and adimer both Whereas that is also deficient in actin bundling, actin in deficient also is that mutant deletion asmaller with observed are results To dimer. induced similar if see of Vt inability the from result may hairpin C-terminal of the deletion with associated defect bundling actin the that suggest results These tions. concentra F-actin athigher intensity in decrease asignificant displays and weight atalower molecular runs band dimer ant To further explore the Vt the To explore dimer further 31 Additionally, we have shown that that shown we have Additionally, Fig. 1B Fig. C5, 1,061–1,066), C5, ). 31,32 32 Δ , a similar decrease in in decrease , asimilar They also proposed proposed also They C5 to form an actin- C5 toan form 32 32 However as our our Howeveras In the Janssen Janssen the In V C2 as for Vt as C2 Δ olume 7Issue 2 31,33 31,33 C2, 1,064– C2, making we have have we C5 vari C5 C2 in in C2 C5 in 30,31 31 31 ------

©2013 Landes Bioscience. Do not distribute. Δ by durotaxis, cell survival or death. survival cell by durotaxis, evidenced as migration ment, including environ their in forces generated locally dependent on are processes cellular Many flow actin. of retrograde not the follow does vinculin Vt, without since, forces for these required be may interaction F-actin/vinculin the the leading edge of cells. edge leading the at events retraction and protrusion during vinculin and of motions F-actin speckle by the displayed of coupling degree to the them correlating and fluctuations, force of measurements through supported ther fur is force transduce and bind F-actin www.landesbioscience.com induce cell morphological changes. morphological cell induce order to in composition adhesion alters stress how mechanical applied to regulate reported been has vinculin forces, external to sense ability its addition In vinculin. including atFAs, proteins through ated medi primarily to be believed is stiffness in changes to sense of cells ability The key factor in regulating how cells respond respond how cells regulating in factor key a to be appears bundling F-actin mediated leading edge of cells. edge leading the within ECM on the traction generate to unable are binding actin in deficient mutants vinculin that shown have studies to stiffen in response to force. response in to stiffen of cells ability the FA and morphology Vt that Δ demonstrated recently also we have addition, In dimer. Vt actin-induced of afunctional mation Y1065 for for Q1066, and critical are residues especially hairpin, C-terminal at FAs. ascaffold as to function of vinculin ability the impact also bundling actin in ciencies defi whether address also and functions, cellular other impact variants deletion bundling-deficientvinculin C-terminal how these examine we will commentary, cellular matrix (ECM). matrix cellular - extra to the of applied force amount the to to FAs corresponds ment of vinculin recruit the since forces, or cell-generated actomyosin either from resulting forces of mechanical for transmission crucial is vinculin that indicates evidence Recent of theF-Actin/Vinculin Interaction C5. 31 These findings suggest that the that suggest findings These The MechanicalRole at Focal Adhesions at 42 The need for vinculin to for vinculin need The 44 However, vinculin-

40,41 C5 is critical for C5 critical is 43 Further, recent recent Further, Furthermore, Furthermore, 31 In this this In 34-36 37-39 ------

spreading, the leading edges of cells are are of cells edges leading the spreading, of cell stages early and migration During forces. external to properly transduce needed are that bundles F-actin culin-mediated vin the but also stimuli, to extracellular to respond necessary are that F-actin and vinculin between connections the only applied. was (FN) fibronectin in coated bead netic amag ofwhen force to pulses responding dling, bun actin in deficient variant vinculin of a expression since force to external larger bundling deficient variant, Vtvariant, Δ deficient bundling larger dimer is observed for Vt Δ Vt for isobserved dimer actin-induced the of amount the in decrease asignificant actin, of presence the In densitometry. ( University)]. Hopkins (John Craig Dr. Susan from [a gift antibody Vt anti-chicken arabbit with blotted and species dimer toobserve panel) gen) (upper (21 band or kDa) monomer Vt the toobserve panel) SDS-PAG on run were samples Crosslinked Vt. against probed =2) and (A/V actin as previously described. Figure 2. *p ≤0.05. *p three independent Statistical experiments. significance was determined using the Student’s t-test. by Vinculin DuringMigration F-Actin BundlesMediated Δ and SpreadingEvents 31 C5 vinculin, prevented cells from from cells prevented C5 vinculin, C These data suggest that it not is that suggest data These Detection of the actin-induced Vt dimer. Crosslinking experiments were conducted conducted were experiments Crosslinking dimer. Vt actin-induced the of Detection ell Adhesion &Migration 31 ( C2 relative toW relative C2 ) Western blots of W of blots A) Western

C5. 31 T Densitometry is the average ±S average isthe Densitometry Vt. Similar results were previously obtained for the the for obtained previously were results Similar Vt. - - - T Vt and Vt Δ Vt and Vt are able to migrate at a higher rate. atahigher able to migrate are cells but the impaired severely is spreading of vinculin, absence the in spreading and mature. cells to move to cells to enable ECM on force the transmits and flowslows actin F-actin, and grins inte active between link the establishes that vinculin and talin including teins, pro cytoskeletal of key recruitment and engagement integrin Following achieved. be spreading and migration effective can components two of these interplay the through Only of adhesions. turnover and formation and polymerization actin in ics - dynam between thebalance by defined of impacting rapid actin polymerization polymerization rapidactin of impacting instead order to in spread adhesions bilize to sta needed is vinculin that suggest data N u PAG B ) Quantification of crosslinked bands by bands crosslinked of ) Quantification E 46 Bis-tris 10% gradient gels (Invitro- gels 10% gradient Bis-tris C2 in the absence and presence of of presence and absence the in C2 In studies examining motility motility examining studies In 45 and allow adhesions to adhesions allow and E M c ombined from from ombined E gels (lower (lower gels 47 These These 221 - - -

©2013 Landes Bioscience. Do not distribute. 222 polymerization and cap filaments. cap and polymerization promote actin can vinculin F-actin, dling to bun addition in that, suggested have studies Recent migration. for effective necessary is that depolymerization and that retains actin binding but possesses but possesses binding actin retains that mutation a vinculin how is does swered multiple defects. possesses likely variant this destabilized, Vt that is likelihood and deletion of this However, size the given migration. during network actin the to organizing taining per functions cellular regulate may action inter F-actin/vinculin the that indicating to the lamellipodia. but FAs, not as tension, such mechanical were under that contractility of high areas to be recruited to vinculin for sufficient was alone domain tail sion of vinculin expres that showed al. et Humphries variants. deletion C-terminal vinculin destabilizing and of large use due to the cells in functions different of these result the to observe hard However, been it has also prevented an invasive phenotype, invasive an prevented also but FAs, denser and not larger played only dis of vinculin domain tail the within 3 2and helix lacking mutant deletion and plated on F on plated and vinculin and Δ vinculin W 0.001 comparing ≤ **p t-test. Student’s the by determined was significance Statistical experiments. independent three from cells ~60 represent Δ expressing Cells ImageJ. Figure 3. One question that remains unan remains that One question

Vinculin C-terminal hairpin deletions alter cell morphology. ( morphology. cell alter deletions hairpin C-terminal Vinculin C2 vinculin toΔ vinculin C2 N for two hours as previously described. previously as hours two for 50 An actin deficient deficient actin An C5 vinculin show significantly more and larger protrusions per cell, relative toW relative cell, per protrusions larger and more significantly show vinculin C5 C5 vinculin. Scale bar is 30 μ is30 bar Scale vinculin. C5 31,48,49 51 ------

W T-, T-, W vitro, in bundling actin in defects similar their vinculin ( WT as of protrusions size and number expressing ( vinculin WT ing express to cells comparison in cells ing for Δ were observed protrusions larger and more significantly ( morphology cell all over and spreading in differences mine deter to quantified was overon time FN significant differences during spreading spreading during differences significant ability of Vin ability fibroblasts (Vin fibroblasts embryo murine null vinculin that shown recently mentioned we have earlier, as its ability to spread on FN. to spread ability its in deficient sitetion (Y1065) slightly is - dimeriza actin-induced proposed the in amutation containing variant vinculin a that shown been It has interactions? to probe vinculin/F-actin used viously pre been have that variants other from differ actin-bundling in defect a specific cells are allowed to spread on FN. to spread allowed are cells when adhesions fewer significantly have Δ C5 vinculin are smaller in cell area and and area cell in smaller are C5 vinculin 31 31 e number of protrusions and ( and protrusions of e number ( m. B Δ

) Δ C2- or Δ C2- T C C2- and Δ and C2- h ell Adhesion &Migration Fig. 3B and C Fig. 3B and Δ C2 vinculin had the same same the had vinculin C2 -/- MEFs expressing either either expressing MEFs -/- A) Vin MEFS) re-expressing re-expressing MEFS) C5 vinculin to spread to spread C5 vinculin Fig. 3B and C Fig. 3B and C5 vinculin express C5 vinculin C5 vinculin display display C5 vinculin -/- Fig. 3 Fig. s were transfected with GFP-tagged W GFP-tagged with transfected s were M E ). Thus, despite Thus, ). F 52 ). Although Although ). Moreover, Moreover,

), cells ), cells 31 C The The ) the relative protrusion area were measured with with measured were area protrusion relative ) the - - - - - tion partners may yet be identified. be yet may partners tion Moreover,interac cells. in new verified not have been interactions proposed these some of identified, been have partners ing of bind anumber While or migration. spreading cell efficient of FA dynamics, regulation vinculin-mediated enabling proteins of additional recruitment tates facili activation vinculin of adhesion, sites At function. promote scaffold its to believed is of vinculin Activation Dimer: Could It Be a Scaffold Too? hairpin of vinculin. C-terminal the in acids amino five vs. two last for roles the differential suggest data These vinculin. WT from as well morphology, as cell overall their in and observed in the presence of the different different of the presence the in observed are that phenotypes the fully, explored be to remains question this While properties? signaling contributeto its and bundles) tension(i.e., F-actin mediate vinculin by activated formed structures the Could raised: is question following tension, the mechanical regulates and senses both that protein scaffolding essential an is vinculin T The F-ActinInducedVinculin vinculin and Δ and vinculin T - C2 vinculin. vinculin. C2 , Δ C2- or Δ or C2- C5 vinculin vinculin C5 R V e olume 7Issue 2 sults shown shown sults 53 N Since Since T I H

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©2013 Landes Bioscience. Do not distribute. expressing either Δ either expressing sion complexes. atadhe accumulated actin show less also and ascaffold, as to act ability their in impaired are deficient mutants bundling actin- the expressing cells that suggest ( of actin amount vinexin- to not recruit only ability their in impaired severely were or untransfected were mutants the expressing However, cells ( vinculin WT expressing cells as of efficiency high as butcomplex not with Arp2/3 the were able to recruit vinculin ( subunit p34-Arc oftion the blot detec by western indicated plex, as dling ( dling actin-bun in deficient mutants vinculin ing site for the Arp2/3 complex ( complex site Arp2/3 for the ing dock P878, the including region line-rich pro the in acids amino with C-terminus the in residues the between of contacts ber anum are protein there full-length the in 1,061–1,066dues However, vinculin. in to bind to resi known are that partners no binding are there Currently function? ascaffolding serve itself dimer the could Furthermore, partners? site for interaction abinding as function also C-terminus the or does dimer actin-induced of the eration gen via bundling topin promote F-actin hair role C-terminal of the primary the Is www.landesbioscience.com dimer itself or residues in the C-terminus C-terminus the in or residues itself dimer actin-induced the whether determined vinexin- recruit were able to efficiently vinculin WT ing contain cells from isolated Adhesions 4 Figure in described tacts con to the close linker proline-rich the in vinculin with interacts also spreading, promotes cell and rearrangements etal cytoskel actin influences that adaptor an vinexin- that suggested been also It has were isolated as previously described. previously as were isolated beads the around formed that complexes FN, with coated beads magnetic with min of 30 time incubation ashort Following adhesions. to integrin-containing partners binding known to recruit ability tor their to moni cells, or untransfected vinculin WT-, either Δ expressing cells from adhesions we isolated mutants, our region? proline-rich to the partners of interaction binding late regu hairpin C-terminal the in residues To assess the scaffolding function of function scaffolding To the assess Fig. 3 Fig. α , but also lacked a significant asignificant lacked , but also ) raise interesting questions. questions. interesting ) raise 31 and the Arp2/3 com Arp2/3 the α and While it remains to be to be it remains While Fig. 5 Fig. C2-, C2-,

Δ ). These results results These ). C5- or lacking orC5- lacking . 18,54,55 Fig. 5 Fig. C2-, C2-, Fig. 4 Fig. Perhaps Perhaps Fig. 5 Fig. ). Cells Cells ). Δ C5 C5 ). α 56 ). ). 21 ------,

ences between the Δ the between ences differ phenotypic examine will studies future Additionally, C-terminus. the within mutations more selective through possibility this explore to further esting inter be it will interactions, these regulate are observed in cells, even though the the though even cells, in observed are between ences differ phenotypic distinct Moreover, Vt dimer. actin-induced of the mation for for the Q1066 crucial to be appear Y1065 residues and vinculin particular, In cells. in and vitro in both bundling F-actin mediated tail vinculin in hairpin C-terminal of the importance the strated demon have studies our from Results vinculin dimer. actin-induced of the formation the impact differences how these and mutations lin the strap, K915 (yellow) in the helix 1/helix 2 loop and P878 (gray) in the proline-rich linker region. linker proline-rich P878 the and in (gray) 2loop 1/helix K915 helix strap, the in the (yellow) T bbon diagram of Vt (PDB ID 1S ID (PDB Vt of diagram bbon Figure 4. h e side chain of of chain e side C

R ell Adhesion &Migration i Conclusions Δ Y 1 C2- and and C2- 065 (red) in the C-terminal hairpin forms hydrogen bonds with D882 (green) in in (green) D882 with bonds hydrogen forms hairpin C-terminal the in (red) 065 C2- and Δ and C2- Δ C5 vinculin C5 vinculin

C5 vincu C5 T 6 ) highlighting interactions of the the of interactions ) highlighting ------and dimerization model. dimerization and following: the explore to further interesting be will it results, Given these complexes. adhesion at of actin accumulation prevent the also mutations deletion hairpin C-terminal of vinexin- recruitment in defective appear mutants deletion C-terminal the of both as function scaffolding vinculin’s to by contributing cells to FAs in partners of binding recruitment play arolethe in may hairpin C-terminal the Furthermore, vitro. in deficiencies actin-bundling same the display variants deletion C-terminal folding function. folding scaf contributeto vinculin’s may dimer or actin-induced hairpin C-terminal C-terminal hairpin for binding. the require that proteins interacting lin (2) Examine if there are other vincu other are there if Examine (2) interaction the F-actin/Vt (1) Refine (3) Further investigate how the how the investigate Further (3) Y 1 065 side chain. chain. side 065 α . The . The 223 - -

©2013 Landes Bioscience. Do not distribute. 224 7. 5. 4. 2. adhe Cell MA. Schwartz AR, JT, Horwitz arsons 1. S.L.C. to 1RO1GM081764 1RO1GM080568 and 1RO1GM029860Health and to K.B. of Institute National the from grants by supported work was This gestions. sug and comments helpful for his Hill) at Chapel Carolina of North (University Peter Thompson M. We thank 6. 3.

M prevent actin accumulation at adhesion complexes. Adhesion complexes were isolated from Vin from isolated were complexes Adhesion complexes. adhesion at accumulation actin prevent Figure 5. beads for 30 min. 30 for beads

M PMID:1618784. 267:12845-50; 1992; Chem J Biol meta-vinculin. to unique exon pair a207-base of splicing alternative by gene a single from derived are meta-vinculin and vinculin Chicken B Z PMID:9580561. 111:1535-44; 1998; Sci JCell vinculin. deleted domain by locomotion cell on effects cells; F9 null in vinculin full-length of reexpression by phenotype X PMID:9486805. 125:327-37; 1998; Development development. onic embry during defects brain and heart in results out X Z nrm2957 http://dx.doi.org/10.1038/ PMID:20729930; 43; 11:633- 2010; Biol Cell Mol Rev Nat tension. lular cel and dynamics cytoskeletal integrating sion: P org/10.1161/01.CIR.95.1.17 http://dx.doi. PMID:8994410; 1997; 95:17-20; Circulation metavinculin. protein cytoskeletal the of deficiency with associated cardiomyopathy Dilated org/10.1016/S0002-9440(10)63364-0 http://dx.doi. PMID:15331426; 165:1033-44; 2004; JPathol Am cardiomyopathy. stress-induced to predisposes gene vinculin the of inactivation Heterozygous al. et ND, KP, Dalton Roos MC, org/10.1016/j.tcb.2006.07.004 http://dx.doi. PMID:16893648; 16:453-60; 2006; Biol Cell Trends vinculin. of regulation and structure F yrne BJ, Kaczorowski YJ, Coutu MD, Craig SW. Craig MD, Coutu YJ, BJ, Kaczorowski yrne s expressing either W either s expressing iegler WH, Liddington RC, Critchley DR. The The DR. Critchley RC, Liddington WH, iegler u W, Coll JL, Adamson ED. Rescue of the mutant mutant the of Rescue ED. Adamson JL, u W, Coll knock Vinculin ED. Adamson H, u W, Baribault emljic-Harpf AE, Ponrartana S, Avalos RT, Jordan RT, Jordan Avalos S, Ponrartana AE, emljic-Harpf aeda M, Holder E, Lowes B, Valent S, Bies RD. RD. Bies S, Valent B, Lowes E, Holder M, aeda Vinculin mutants deficient in F-actin bundling also alter vinculin’s scaffold function and function scaffold vinculin’s alter also bundling F-actin in deficient mutants Vinculin Acknowledgments References T - , Δ C2-, Δ C2-, C5 vinculin or untransfected, after incubation with F with incubation after untransfected, or vinculin C5 - - - - - 15. 14. 12. 10. 8. 16. 13. 11. 9.

http://dx.doi.org/10.1016/0006-291X(87)90564-X PMID:3113423; 1987; 146:554-60; Commun Res Biophys Biochem alpha-actinin. with vinculin of tion org/10.1083/jcb.142.3.847 http://dx.doi. PMID:9700171; 142:847-57; 1998; Biol JCell cells. epithelial in complex junctional apical the organize to functions interaction vinculin alpha-Catenin- T, al. et Uemura K, Fujimoto A, org/10.1038/nature02610 PMID:15195105; http://dx.doi. 430:583-6; 2004; Nature adhesion. cell of sites at activation vinculin for basis Structural al. et L, GW, Jennings Cadwell org/10.1073/pnas.92.20.9161 http://dx.doi. 92:9161-5; PMID:7568093; A 1995; US Sci Acad Natl Proc locomotion. and sion, adhe morphology, cell changes cells stem embryonic and F9 in genes vinculin of disruption Targeted al. et RG, Oshima H, Baribault JL, Fernández http://dx.doi.org/10.1093/emboj/16.10.2717 J1997; PMID:9184218; 16:2717-29; EMBO invasin. IpaA Shigella the and vinculin between association by cells epithelial into entry bacterial of Modulation org/10.1074/jbc.M508060200 http://dx.doi. PMID:16135522; 280:37217-24; 2005; Chem J Biol rod. talin the within sites binding vinculin multiple for identification sequence sensus con and Mapping al. et DR, Critchley GC, Roberts jbc.272.51.32448 http://dx.doi.org/10.1074/ PMID:9405455; 53; 1997; 272:32448- Chem JBiol complex. adhesion E-cadherin the with associated is Vinculin DL. H B jcb.200308011 81; PMID:15138291; http://dx.doi.org/10.1083/ 165:371- 2004; Biol J Cell motility. and survival control to ERK regulates interactions illin-FAK pax of modulation Vinculin KM. Hahn S, Junger S C http://dx.doi.org/10.1161/hc0402.102930 PMID:11815424; 105:431-7; 2002; thy. Circulation cardiomyopa dilated in interaction actin alter tions muta Metavinculin BM. MT, Jockusch Keating S, O T G W W ubauste MC, Pertz O, Adamson ED, Turner CE, Turner CE, ED, O, Adamson Pertz MC, ubauste ran Van Nhieu G, Ben-Ze’ev A, Sansonetti PJ. Sansonetti A, Ben-Ze’ev G, Van Nhieu ran akolitsa C, Cohen DM, Bankston LA, Bobkov AA, AA, Bobkov LA, Bankston DM, Cohen C, akolitsa oll JL, Ben-Ze’ev A, Ezzell RM, Rodríguez Rodríguez RM, Ezzell A, Ben-Ze’ev JL, oll ingras AR, Ziegler WH, Frank R, Barsukov IL, IL, Barsukov R, Frank WH, Ziegler AR, ingras lson TM, Illenberger S, Kishimoto NY, Huttelmaier Huttelmaier NY, Kishimoto S, Illenberger TM, lson azan RB, Kang L, Roe S, Borgen PI, Rimm Rimm PI, Borgen S, Roe L, Kang RB, azan achsstock DH, Wilkins JA, Lin S. Specific interac Specific S. Lin JA, Wilkins DH, achsstock Y, Nagafuchi Imamura N, Uchida M, atabe-Uchida C ell Adhesion &Migration

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------19. 27. 26. 25. 20. 18. K, Takahashi A, Satoh H, Nakanishi K, andai 17. 30. 29. 28. 24. 23. 22. 21.

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