DOCSLIB.ORG

Rab Gtpases at a Glance Samantha L

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

Author Correction Rab GTPases at a glance Samantha L. Schwartz, Canhong Cao, Olena Pylypenko, Alexey Rak and Angela Wandinger-Ness Journal of Cell Science 121, 246 (2008) doi:10.1242/jcs.03495 There was an error published in J. Cell Sci. 120, 3905-3910. The authors regret that in Table 1 under ‘(13) Rab1, Rab3, Rab8, Rab10, Rab12, Rab13, Rab15, Rab35, Rab40’, Rab35 was incorrectly referred to as an uncharacterized member of the Rab family. The correct details are given below. Rab Localization Function Reference Rab35 Plasma membrane, endosomes, intercellular bridge Rapid endocytic recycling and cytokinesis Kouranti et al., 2006 Cell Science at a Glance 3905 Rab GTPases at a brain) GTPases were first identified as associate with regulatory molecules and evolutionarily conserved, essential downstream effectors to exert their glance regulators of membrane trafficking functions (Chen et al., 2003; Pereira- Samantha L. Schwartz1, Canhong (Salminen and Novick, 1987; Schmitt et Leal and Seabra, 2000; Pfeffer, 2005). Cao1, Olena Pylypenko2 Alexey al., 1986; Touchot et al., 1987). The Rak2,3 and Angela Wandinger- proteins are members of the wider Ras Ness1,* superfamily of GTPases (Wennerberg et Rab GTPases regulate membrane al., 2005). Over 70 human Rab and Rab- trafficking, cell growth and 1Department of Pathology MSC08-4640, University of New Mexico, 2325 Camino de Salud NE, like members of the Ras superfamily differentiation CRF225, Albuquerque, NM 87131, USA have been identified (Colicelli, 2004; Characterization of around half of the 2Departments of Structural Biology and Physical Stenmark and Olkkonen, 2001), and the known Rab GTPases has revealed the Biochemistry, Max Planck Institute for Molecular Physiology, 44227 Dortmund, Germany functions of 36 Rab GTPases have been extraordinary complexity of membrane 3Sanofi-Aventis, Centre de Recherche Paris,13, delineated. Rab GTPases are molecular trafficking circuits and shows that Rab Quai Jules Guesde–BP 14, 94403 Vitry sur Seine Cedex, France switches, cycling between active and GTPases are also essential for signaling *Author for correspondence (e-mail: inactive states and serving as scaffolds to and the control of cell proliferation [email protected]) integrate both membrane trafficking and and differentiation. Rab proteins are intracellular signaling in a temporally present on all compartments of the Journal of Cell Science 120, 3905-3910 Published by The Company of Biologists 2007 and spatially sensitive manner (Bucci endomembrane system (endoplasmic doi:10.1242/jcs.015909 and Chiariello, 2006; Zerial and reticulum, Golgi, endosomes, McBride, 2001). Despite the small sizes lysosomes), the nucleus, the plasma It has been two decades since the yeast of Rab proteins (20-25 kDa), structural membrane (including cell junctions and Ypt1 and Sec4 proteins and the analyses reveal they have multiple focal adhesions), mitochondria and mammalian Rab (Ras-related proteins in interaction surfaces through which they centrioles. In addition, they help regulate Rab GTPases at a Glance Samantha L. Schwartz, Canhong Cao, Olena Pylypenko, Alexey Rak and Angela Wandinger-Ness Rab structures Rab7-GTP Cilium jcs.biologists.org MTM1CS Rab5Q79L VPS34 IFT Rab7-GDP RAB7N1251I Rab7 Journal of Cell Science Rab12 Caveolae Rab7-GTP-RILP dimer (PDB ID 1YHN) Rab7-GDP-REP1 (PDB ID 1VGO) Apical membrane Rab23 Vesicle transport Secretory Rab8 CCV granule Centrioles Rab15 Donor membrane Melanosome Rab5a-c Caveosome Coat Rab4 Rab11a M6PR Rab13 Budding GDF TIP47 (fast) GDP Constitutive (slow) R GDP R ab ab M6P ab Rab4a GTP endocytosis R Early sorting Recycling REP endosome Rab17 Rab8b GEF GTP endosome GGT Rab GDP Rab27 Rab11a GDP Motor Regulated protein exocytosis Rab38 Rab22b Rab25 Cytoskeletal Rab32 MPC R Rab11a GDP transport R ab Rab27a,b Rab17 ab GTP Rab22a Rab7 SNARE Rab26 GDI Rab14 Basolateral Rab37 Tethering factor Rab6a membrane Rab3a-d GTP Rab9 GAP Late endosome Rab Rab11b Rab6a,b Pi Rab33b Rab14 GDP Cytoskeletal Rab22a ab Phagocytosis R filaments Rab7b Docking/fusion GDI Rab8 Endoplasmic reticulum Golgi Rab2 Lyososome Phagosome Acceptor membrane complex Signaling Rab1 EGF signaling Rab34 Macropinosome EGF Rab10 Rab23 Macropinocytosis NuRD Rab5 APPL1 APPL1 Rab34 GTP MeCP1 Autophagosome Basolateral Early Nucleus Sequestration of APPL1 Chromatin remodeling and changes Rab18 Rab5 membrane endosome on endosomes in gene expression in nucleus Rab24 GDP Lipid Rab3ab332 Relocation to cytoplasm Rab5 droplets Rab21 Metabolic signaling Insulin/AKT Ran MitochondriMitochondria Rab2 Rab8A Rab Rab8B GAPTbc1d4 Rab Rab10 GTP Tbc1d1 Rab14 GTP Endocytosis Focal adhesion PO4 Increased fusion of Glut4 Rab inactivation by Rab vesicles with plasma membrane GAP on cytoplasmic GAP GDP Glut4 storage vessels GAP inactivation Abbreviations: CCV, clathrin-coated vesicle; GAP, GTPase activating protein; GDF, GDI dissociation factor; GDI, GDP dissociation inhibitor; GGT, geranylgeranyltransferase; GDP, guanosine diphosphate; GEF, guanine nucleotide exchange factor; GTP, guanosine triphosphate; IFT, intraflagellar transport; M6P, mannose 6-phosphate; M6PR, cation-independent mannose 6-phosphate receptor; MPC, motor protein complex; Rab, Ras-related in brain; REP, Rab escort protein; RILP, Rab interacting lysosomal protein; SNARE, soluble NSF attachment receptor; TIP47, tail-interacting protein 47kD. © Journal of Cell Science 2007 (120, pp. 3905-3910) (See poster insert) 3906 Journal of Cell Science 120 (22) a vast array of basic cellular functions – destinations (colored lines on the poster junctional proteins and integrins and by from macromolecular homeostasis to denote such domains; the micrograph defining epithelial transport circuits to growth control. illustrates alternating Rab7 and Rab5 cilia (connecting with intraflagellar domains on dilated early endosomes) transport, IFT), the apical (AM) and Rab proteins are best known for their (Barbero et al., 2002; Vitale et al., 1998; basolateral (BM) membranes, and apical essential roles in exocytic and endocytic Vonderheit and Helenius, 2005). In this recycling endosomes (AREs). They thus membrane trafficking, which encompass way, Rab GTPases regulate plasma play major regulatory roles maintaining the constitutive and regulated secretory membrane delivery, organelle biogenesis compartment identity, regulating cargo routes, endocytosis via caveolae or and degradative pathways (lysosomal and delivery, controlling protein and lipid clathrin-coated vesicles (CCVs), autophagic). They also contribute to cell- storage/degradation and modulating micropinocytosis and phagocytosis. They type-specific functions, such as regulated specialized trafficking functions. control anterograde and retrograde secretion (secretory granules/lysosomes trafficking between compartments to in endocrine and exocrine cells), synaptic Rab proteins are increasingly found coordinate cargo delivery and membrane transmission [synaptic vesicles (SVs) in downstream of signaling cascades and recycling and also subcompartmentalize neurons] and phagocytosis (in can impact gene expression and growth organelles by organizing specific macrophages and dendritic cells). In control. Rab5, for example, is implicated membrane domains that function in epithelia, Rab GTPases help generate in EGF signaling and thought to trafficking of cargo to different polarity by regulating the trafficking of sequester APPL1, an adaptor protein Table 1. The Rab family Rab Localization Function References (1) Rab23 Rab23 Plasma membrane and endosomes Trafficking of sonic hedgehog signaling components; Evans et al., 2003 embryogenesis; ciliary trafficking (2) Rab29, Rab32, Rab38, Rab7L1 Rab32 Perinuclear vesicles; mitochondria Post-Golgi trafficking of melanogenic enzymes; binds Alto et al., 2002; Wasmeier et al., 2006 PKA and regulates mitochondrial dynamics Rab38 Tyrosinase-positive vesicles Post-Golgi biogenesis of melanosomes Wasmeier et al., 2006 (3) RabL2, RabL3, RabL5 (4) Ran Ran Nucleus, cytoplasm Nucleocytoplasmic transport Joseph, 2006 (5) Rab7, Rab7b, Rab9 Journal of Cell Science Rab7, Rab7b Late endosomes, lysosomes Transport from early to late endosomes; lysosome Feng et al., 2001; Bucci et al., 2000; Wang biogenesis; b) transport to lysosomes, TLR4 et al., 2007 signaling Rab9a,b,c Late endosomes Lysosomal enzyme and cholesterol trafficking; late Ganley et al., 2004; Lombardi et al., 1993; endosome to trans-Golgi transport Narita et al., 2005 (6) Rab28, RabL4 (7) Rab34, Rab36 Rab34 Cell surface membrane ruffles, Regulation of spatial distribution of lysosomes; Colucci et al., 2005; Sun and Endo, 2005; lysosomes Formation of macropinosomes Sun et al., 2003; Wu et al., 2005 (8) Rab6, Rab41 Rab6a,a′,b,c (a,a′,b) Golgi, (b) ERGIC-53-positive Retrograde transport; (a,b) Golgi to ER and intra- Jiang and Storrie, 2005; Martinez et al., vesicles and neuronal cell specific Golgi tranport,Golgi stress response (a′) endosome to 1994; Del Nery et al., 2006; Opdam et al., Golgi transport; (c) multi-drug resistance regulation 2000; Shan et al., 2000 (9) Rab5, Rab17, Rab20, Rab21, Rab22a/Rab31, Rab24 Rab5a,b,c Clathrin coated vesicles, caveosomes, Endocytosis, early endosome fusion, caveolar vesicle Pelkmans et al., 2004; Barbieri et al., 2000; and early endosomes. targeting to early endosomes; (a) EGF receptor Arnett et al., 2004; Bucci et al., 1995 activation; (b) neuroprotection Rab17 Epithelial specific; apical recycling Transport through apical recycling endosomes; Zacchi et al., 1998 endosome polarized sorting Rab20 Epithelial specific; kidney
Recommended publications
  • Anti-Rab11 Antibody (ARG41900)

    Anti-Rab11 Antibody (ARG41900)

    Product datasheet [email protected] ARG41900 Package: 100 μg anti-Rab11 antibody Store at: -20°C Summary Product Description Goat Polyclonal antibody recognizes Rab11 Tested Reactivity Hu, Ms, Rat, Dog, Mk Tested Application IHC-Fr, IHC-P, WB Host Goat Clonality Polyclonal Isotype IgG Target Name Rab11 Antigen Species Mouse Immunogen Purified recombinant peptides within aa. 110 to the C-terminus of Mouse Rab11a, Rab11b and Rab11c (Rab25). Conjugation Un-conjugated Alternate Names RAB11A: Rab-11; Ras-related protein Rab-11A; YL8 RAB11B: GTP-binding protein YPT3; H-YPT3; Ras-related protein Rab-11B RAB25: RAB11C; CATX-8; Ras-related protein Rab-25 Application Instructions Application table Application Dilution IHC-Fr 1:100 - 1:400 IHC-P 1:100 - 1:400 WB 1:250 - 1:2000 Application Note IHC-P: Antigen Retrieval: Heat mediation was recommended. * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. Positive Control Hepa cell lysate Calculated Mw 24 kDa Observed Size ~ 26 kDa Properties Form Liquid Purification Affinity purification with immunogen. Buffer PBS, 0.05% Sodium azide and 20% Glycerol. Preservative 0.05% Sodium azide www.arigobio.com 1/3 Stabilizer 20% Glycerol Concentration 3 mg/ml Storage instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use. Note For laboratory research only, not for drug, diagnostic or other use.
  • Uwaterloo Latex Thesis Template

    Uwaterloo Latex Thesis Template

    New Algorithms for Predicting Conformational Polymorphism and Inferring Direct Couplings for Side Chains of Proteins by Laleh Soltan Ghoraie A thesis presented to the University of Waterloo in fulfillment of the thesis requirement for the degree of Doctor of Philosophy in Computer Science Waterloo, Ontario, Canada, 2015 c Laleh Soltan Ghoraie 2015 I hereby declare that I am the sole author of this thesis. This is a true copy of the thesis, including any required final revisions, as accepted by my examiners. I understand that my thesis may be made electronically available to the public. ii Abstract Protein crystals populate diverse conformational ensembles. Despite much evidence that there is widespread conformational polymorphism in protein side chains, most of the x- ray crystallography data are modelled by single conformations in the Protein Data Bank. The ability to extract or to predict these conformational polymorphisms is of crucial im- portance, as it facilitates deeper understanding of protein dynamics and functionality. This dissertation describes a computational strategy capable of predicting side-chain poly- morphisms. The applied approach extends a particular class of algorithms for side-chain prediction by modelling the side-chain dihedral angles more appropriately as continuous rather than discrete variables. Employing a new inferential technique known as particle belief propagation (PBP), we predict residue-specific distributions that encode informa- tion about side-chain polymorphisms. The predicted polymorphisms are in relatively close agreement with results from a state-of-the-art approach based on x-ray crystallography data. This approach characterizes the conformational polymorphisms of side chains us- ing electron density information, and has successfully discovered previously unmodelled conformations.
  • Mechanical Forces Induce an Asthma Gene Signature in Healthy Airway Epithelial Cells Ayşe Kılıç1,10, Asher Ameli1,2,10, Jin-Ah Park3,10, Alvin T

    Mechanical Forces Induce an Asthma Gene Signature in Healthy Airway Epithelial Cells Ayşe Kılıç1,10, Asher Ameli1,2,10, Jin-Ah Park3,10, Alvin T

    www.nature.com/scientificreports OPEN Mechanical forces induce an asthma gene signature in healthy airway epithelial cells Ayşe Kılıç1,10, Asher Ameli1,2,10, Jin-Ah Park3,10, Alvin T. Kho4, Kelan Tantisira1, Marc Santolini 1,5, Feixiong Cheng6,7,8, Jennifer A. Mitchel3, Maureen McGill3, Michael J. O’Sullivan3, Margherita De Marzio1,3, Amitabh Sharma1, Scott H. Randell9, Jefrey M. Drazen3, Jefrey J. Fredberg3 & Scott T. Weiss1,3* Bronchospasm compresses the bronchial epithelium, and this compressive stress has been implicated in asthma pathogenesis. However, the molecular mechanisms by which this compressive stress alters pathways relevant to disease are not well understood. Using air-liquid interface cultures of primary human bronchial epithelial cells derived from non-asthmatic donors and asthmatic donors, we applied a compressive stress and then used a network approach to map resulting changes in the molecular interactome. In cells from non-asthmatic donors, compression by itself was sufcient to induce infammatory, late repair, and fbrotic pathways. Remarkably, this molecular profle of non-asthmatic cells after compression recapitulated the profle of asthmatic cells before compression. Together, these results show that even in the absence of any infammatory stimulus, mechanical compression alone is sufcient to induce an asthma-like molecular signature. Bronchial epithelial cells (BECs) form a physical barrier that protects pulmonary airways from inhaled irritants and invading pathogens1,2. Moreover, environmental stimuli such as allergens, pollutants and viruses can induce constriction of the airways3 and thereby expose the bronchial epithelium to compressive mechanical stress. In BECs, this compressive stress induces structural, biophysical, as well as molecular changes4,5, that interact with nearby mesenchyme6 to cause epithelial layer unjamming1, shedding of soluble factors, production of matrix proteins, and activation matrix modifying enzymes, which then act to coordinate infammatory and remodeling processes4,7–10.
  • Comprehensive Analyses of 723 Transcriptomes Enhance Genetic and Biological Interpretations for Complex Traits in Cattle

    Comprehensive Analyses of 723 Transcriptomes Enhance Genetic and Biological Interpretations for Complex Traits in Cattle

    Downloaded from genome.cshlp.org on October 3, 2021 - Published by Cold Spring Harbor Laboratory Press Resource Comprehensive analyses of 723 transcriptomes enhance genetic and biological interpretations for complex traits in cattle Lingzhao Fang,1,2,3,4,8 Wentao Cai,2,5,8 Shuli Liu,1,5,8 Oriol Canela-Xandri,3,4,8 Yahui Gao,1,2 Jicai Jiang,2 Konrad Rawlik,3 Bingjie Li,1 Steven G. Schroeder,1 Benjamin D. Rosen,1 Cong-jun Li,1 Tad S. Sonstegard,6 Leeson J. Alexander,7 Curtis P. Van Tassell,1 Paul M. VanRaden,1 John B. Cole,1 Ying Yu,5 Shengli Zhang,5 Albert Tenesa,3,4 Li Ma,2 and George E. Liu1 1Animal Genomics and Improvement Laboratory, Henry A. Wallace Beltsville Agricultural Research Center, Agricultural Research Service, USDA, Beltsville, Maryland 20705, USA; 2Department of Animal and Avian Sciences, University of Maryland, College Park, Maryland 20742, USA; 3The Roslin Institute, Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Midlothian EH25 9RG, United Kingdom; 4Medical Research Council Human Genetics Unit at the Medical Research Council Institute of Genetics and Molecular Medicine, The University of Edinburgh, Edinburgh EH4 2XU, United Kingdom; 5College of Animal Science and Technology, China Agricultural University, Beijing 100193, China; 6Acceligen, Eagan, Minnesota 55121, USA; 7Fort Keogh Livestock and Range Research Laboratory, Agricultural Research Service, USDA, Miles City, Montana 59301, USA By uniformly analyzing 723 RNA-seq data from 91 tissues and cell types, we built a comprehensive gene atlas and studied tissue specificity of genes in cattle. We demonstrated that tissue-specific genes significantly reflected the tissue-relevant biol- ogy, showing distinct promoter methylation and evolution patterns (e.g., brain-specific genes evolve slowest, whereas testis- specific genes evolve fastest).
  • Exosomes from Nischarin-Expressing Cells Reduce Breast Cancer Cell Motility and Tumor Growth

    Exosomes from Nischarin-Expressing Cells Reduce Breast Cancer Cell Motility and Tumor Growth

    Author Manuscript Published OnlineFirst on January 11, 2019; DOI: 10.1158/0008-5472.CAN-18-0842 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Exosomes from Nischarin-Expressing Cells Reduce Breast Cancer Cell Motility and tumor growth Mazvita Maziveyi1,7, Shengli Dong1, Somesh Baranwal2, Ali Mehrnezhad3, Rajamani Rathinam4, Thomas M. Huckaba5, Donald E. Mercante6, Kidong Park3, Suresh K. Alahari1 1Department of Biochemistry and Molecular Biology, LSUHSC School of Medicine, New Orleans, LA, USA 2Center of Biochemistry and Microbial Science, Central University of Punjab, Bathinda-151001, India 3Department of Electrical Engineering and Computer Engineering, Louisiana State University, Baton Rouge, LA, USA 4Wayne State University, Detroit, MI, USA 5Department of Biology, Xavier University of Louisiana, New Orleans, LA, USA 6School of Public Health, LSUHSC School of Medicine, New Orleans, LA, USA 7Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas *Corresponding author; Suresh K. Alahari, PhD; Fred G. Brazda Professor of Biochemistry, LSUHSC School of Medicine, New Orleans, LA 70112, USA; Tel: 504-568-4734 [email protected] Running Title: Nischarin regulates exosome production Conflicts of Interest No potential conflicts of interest were disclosed. Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 2019 American Association for Cancer Research. Author Manuscript Published OnlineFirst on January 11, 2019; DOI: 10.1158/0008-5472.CAN-18-0842 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Abstract: Exosomes are small extracellular microvesicles that are secreted by cells when intracellular multivesicular bodies (MVB) fuse with the plasma membrane.
  • Disruption of Adipose Rab10-Dependent Insulin

    Disruption of Adipose Rab10-Dependent Insulin

    Page 1 of 35 Diabetes Vazirani et al. Disruption of Adipose Rab10-Dependent Insulin Signaling Causes Hepatic Insulin Resistance Reema P. Vazirani1, Akanksha Verma2, L. Amanda Sadacca1, Melanie S. Buckman3, Belen Picatoste1, Muheeb Beg1, Christopher Torsitano1, Joanne H. Bruno1, Rajesh T. Patel1, Kotryna Simonyte4, Joao P. Camporez5, Gabriela Moreira5, Domenick J. Falcone6, Domenico Accili7, Olivier Elemento2, Gerald I. Shulman6,8, Barbara B. Kahn4 and Timothy E. McGraw1,3* 1Department of Biochemistry, Weill Cornell Medical College 2Department of Physiology and Biophysics, Weill Cornell Medical College 3Department of Cardiothoracic Surgery, Weill Cornell Medical College 4Beth Israel Deaconess Medical Center, Harvard Medical School 5Departments of Internal Medicine and Cellular & Molecular Physiology, Yale University 6Department of Pathology, Weill Cornell Medical College 7Department of Medicine and Berrie Diabetes Center, Columbia University 8Howard Hughes Medical Institute, Yale University *Contact: Timothy E. McGraw, PhD 1300 York Ave New York, NY 10065 212-746-4982 [email protected] The authors have declared that no conflict of interest exists. 1 Diabetes Publish Ahead of Print, published online March 25, 2016 Diabetes Page 2 of 35 Vazirani et al. Abstract Insulin controls glucose uptake into adipose and muscle cells by regulating the amount of the Glut4 glucose transporter in the plasma membrane. The effect of insulin is to promote translocation of intracellular Glut4 to the plasma membrane. The small Rab GTPase Rab10 is required for insulin-stimulated Glut4 translocation in cultured 3T3-L1 adipocytes. Here we demonstrate that both insulin-stimulated glucose uptake and Glut4 translocation to the plasma membrane are reduced by about half in adipocytes from adipose-specific Rab10 knockout mice.
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus

    A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus

    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
  • Tracking Melanosomes Inside a Cell to Study Molecular Motors and Their Interaction

    Tracking Melanosomes Inside a Cell to Study Molecular Motors and Their Interaction

    Tracking melanosomes inside a cell to study molecular motors and their interaction Comert Kural*, Anna S. Serpinskaya†, Ying-Hao Chou†, Robert D. Goldman†, Vladimir I. Gelfand†‡, and Paul R. Selvin*§¶ *Center for Biophysics and Computational Biology and §Department of Physics, University of Illinois at Urbana–Champaign, Urbana, IL 61801; and †Department of Cell and Molecular Biology, Northwestern University School of Medicine, Chicago, IL 60611 Communicated by Gordon A. Baym, University of Illinois at Urbana–Champaign, Urbana, IL, January 9, 2007 (received for review June 4, 2006) Cells known as melanophores contain melanosomes, which are membrane organelles filled with melanin, a dark, nonfluorescent pigment. Melanophores aggregate or disperse their melanosomes when the host needs to change its color in response to the environment (e.g., camouflage or social interactions). Melanosome transport in cultured Xenopus melanophores is mediated by my- osin V, heterotrimeric kinesin-2, and cytoplasmic dynein. Here, we describe a technique for tracking individual motors of each type, both individually and in their interaction, with high spatial (Ϸ2 nm) and temporal (Ϸ1 msec) localization accuracy. This method enabled us to observe (i) stepwise movement of kinesin-2 with an average step size of 8 nm; (ii) smoother melanosome transport (with fewer pauses), in the absence of intermediate filaments (IFs); and (iii) motors of actin filaments and microtubules working on the same cargo nearly simultaneously, indicating that a diffusive step is not needed between the two systems of transport. In concert with our previous report, our results also show that dynein-driven retro- grade movement occurs in 8-nm steps. Furthermore, previous studies have shown that melanosomes carried by myosin V move 35 nm in a stepwise fashion in which the step rise-times can be as long as 80 msec.
  • High-Throughput Discovery of Novel Developmental Phenotypes

    High-Throughput Discovery of Novel Developmental Phenotypes

    High-throughput discovery of novel developmental phenotypes The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Dickinson, M. E., A. M. Flenniken, X. Ji, L. Teboul, M. D. Wong, J. K. White, T. F. Meehan, et al. 2016. “High-throughput discovery of novel developmental phenotypes.” Nature 537 (7621): 508-514. doi:10.1038/nature19356. http://dx.doi.org/10.1038/nature19356. Published Version doi:10.1038/nature19356 Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:32071918 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use#LAA HHS Public Access Author manuscript Author ManuscriptAuthor Manuscript Author Nature. Manuscript Author Author manuscript; Manuscript Author available in PMC 2017 March 14. Published in final edited form as: Nature. 2016 September 22; 537(7621): 508–514. doi:10.1038/nature19356. High-throughput discovery of novel developmental phenotypes A full list of authors and affiliations appears at the end of the article. Abstract Approximately one third of all mammalian genes are essential for life. Phenotypes resulting from mouse knockouts of these genes have provided tremendous insight into gene function and congenital disorders. As part of the International Mouse Phenotyping Consortium effort to generate and phenotypically characterize 5000 knockout mouse lines, we have identified 410 Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms #Corresponding author: [email protected].
  • Role of Cdc42 in Melanosome Transfer 1443 Approximate Ratio of 1:1 in KGM

    Role of Cdc42 in Melanosome Transfer 1443 Approximate Ratio of 1:1 in KGM

    Research Article 1441 Filopodia are conduits for melanosome transfer to keratinocytes Glynis Scott, Sonya Leopardi, Stacey Printup and Brian C. Madden Department of Dermatology, University of Rochester School of Medicine and Dentistry, Rochester, NY, USA Author for correspondence (e-mail: [email protected]) Accepted 4 January 2002 Journal of Cell Science 115, 1441-1451 (2002) © The Company of Biologists Ltd Summary Melanosomes are specialized melanin-synthesizing cultured with keratinocytes induced a highly dendritic organelles critical for photoprotection in the skin. phenotype with extensive contacts between melanocytes Melanosome transfer to keratinocytes, which involves and keratinocytes through filopodia, many of which whole organelle donation to another cell, is a unique contained melanosomes. These results suggest a unique role biological process and is poorly understood. Time-lapse for filopodia in organelle transport and, in combination digital movies and electron microscopy show that filopodia with our previous work showing the presence of SNARE from melanocyte dendrites serve as conduits for proteins and rab3a on melanosomes, suggest a novel model melanosome transfer to keratinocytes. Cdc42, a small system for melanosome transfer to keratinocytes. GTP-binding protein, is known to mediate filopodia formation. Melanosome-enriched fractions isolated from Movies available on-line human melanocytes expressed the Cdc42 effector proteins PAK1 and N-WASP by western blotting. Expression of Key words: Melanosome, Melanocyte, Cdc42, Filopodia, constitutively active Cdc42 (Cdc42V12) in melanocytes co- Keratinocyte Introduction microscopy of cultured cells, which allowed direct Melanosomes are organelles unique to melanocytes that visualization of melanosome movement and modifiers of function in the synthesis of melanin, a complex pigment actin, microtubules and their motor proteins.
  • Four-Dimensional Live Imaging of Apical Biosynthetic Trafficking Reveals a Post-Golgi Sorting Role of Apical Endosomal Intermediates

    Four-Dimensional Live Imaging of Apical Biosynthetic Trafficking Reveals a Post-Golgi Sorting Role of Apical Endosomal Intermediates

    Four-dimensional live imaging of apical biosynthetic trafficking reveals a post-Golgi sorting role of apical endosomal intermediates Roland Thuenauera,b,1,2, Ya-Chu Hsua, Jose Maria Carvajal-Gonzaleza,3, Sylvie Debordea,4, Jen-Zen Chuanga, Winfried Römerc,d, Alois Sonnleitnerb, Enrique Rodriguez-Boulana,5, and Ching-Hwa Sunga,5 aMargaret M. Dyson Vision Research Institute, Weill Medical College of Cornell University, New York, NY 10065; bCenter for Advanced Bioanalysis Linz, 4020 Linz, Austria; and cInstitute of Biology II, and dBIOSS Centre for Biological Signalling Studies, Albert-Ludwigs-University Freiburg, 79104 Freiburg, Germany Edited by Keith E. Mostov, University of California School of Medicine, San Francisco, CA, and accepted by the Editorial Board January 17, 2014 (received for review March 11, 2013) Emerging data suggest that in polarized epithelial cells newly is an important regulator of biological processes that require synthesized apical and basolateral plasma membrane proteins apical trafficking, e.g., lumen formation during epithelial tubu- traffic through different endosomal compartments en route to the logenesis (11), apical secretion of discoidal/fusiform vesicles in respective cell surface. However, direct evidence for trans-endo- bladder umbrella cells (12), and apical microvillus morphogenesis somal pathways of plasma membrane proteins is still missing and and rhodopsin localization in fly photoreceptors (13). However, the mechanisms involved are poorly understood. Here, we imaged despite the physiological importance of trans-endosomal traf- the entire biosynthetic route of rhodopsin-GFP, an apical marker in ficking, the underlying mechanisms remain largely unclear. epithelial cells, synchronized through recombinant conditional ag- Previous studies on trans-endosomal trafficking in polarized gregation domains, in live Madin-Darby canine kidney cells using epithelial cells have relied on pulse chase/cell fractionation pro- spinning disk confocal microscopy.
  • ADP-Ribosylation Factor, a Small GTP-Binding Protein, Is Required for Binding of the Coatomer Protein Fl-COP to Golgi Membranes JULIE G

    ADP-Ribosylation Factor, a Small GTP-Binding Protein, Is Required for Binding of the Coatomer Protein Fl-COP to Golgi Membranes JULIE G

    Proc. Natl. Acad. Sci. USA Vol. 89, pp. 6408-6412, July 1992 Biochemistry ADP-ribosylation factor, a small GTP-binding protein, is required for binding of the coatomer protein fl-COP to Golgi membranes JULIE G. DONALDSON*, DAN CASSEL*t, RICHARD A. KAHN*, AND RICHARD D. KLAUSNER* *Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, and tLaboratory of Biological Chemistry, Division of Cancer Treatment, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 Communicated by Marc Kirschner, April 20, 1992 (receivedfor review February 11, 1992) ABSTRACT The coatomer is a cytosolic protein complex localized to the Golgi complex, although their functions have that reversibly associates with Golgi membranes and is Impli- not been defined. Distinct among these proteins is the ADP- cated in modulating Golgi membrane transport. The associa- ribosylation factor (ARF), originally identified as a cofactor tion of 13-COP, a component of coatomer, with Golgi mem- required for in vitro cholera toxin-catalyzed ADP- branes is enhanced by guanosine 5'-[v-thioltriphosphate ribosylation of the a subunit of the trimeric GTP-binding (GTP[yS]), a nonhydrolyzable analogue of GTP, and by a protein G, (G,.) (19). ARF is an abundant cytosolic protein mixture of aluminum and fluoride ions (Al/F). Here we show that reversibly associates with Golgi membranes (20, 21). that the ADP-ribosylation factor (ARF) is required for the ARF has been shown to be present on Golgi coated vesicles binding of (-COP. Thus, 13-COP contained in a coatomer generated in the presence of GTP[yS], but it is not a com- fraction that has been resolved from ARF does not bind to Golgi ponent of the cytosolic coatomer (22).