Vps33b regulates Vwf‐positive vesicular trafficking in megakaryocytes

Mutations of vacuolar protein sorting‐associated protein 33b (VPS33B) cause arthrogryposis, renal dysfunction, and cholestasis syndrome, and a lack of platelet α‐granules in the affected patients. Conditional Vps33b knockout mice were developed to investigate the function(s) of Vps33b in platelet α‐granule formation. We found that early embryonic deletion of Vps33b was lethal. PF4‐Cre‐driven megakaryocyte‐targeted Vps33b gene deletion greatly diminished Vps33b expression in platelets, but had no effect on platelet α‐granule formation and protein content. Tamoxifen‐induced, haematopoietic stem cell (HSC)‐specific Vps33b deletion completely depleted Vps33b in platelets, caused the absence of α‐granules, and increased the number of vacuoles in platelets and megakaryocytes. VPS33B association with VIPAS39, α‐tubulin, and SEC22B was identified by co‐immunoprecipitation, mass spectra, and immunoblotting in human embryonic kidney 293T (HEK293T) cells. Also, pull‐down experiments revealed that VIPAS39 bound to intact VPS33B; in contrast, α‐tubulin and SEC22B separately interacted with the sec1‐like domains of VPS33B. Vps33b deficiency in megakaryocytes disturbs the redistribution of Vipas39 and Sec22b to proplatelets, and interrupted the co‐localization of Sec22b with Vwf‐positive vesicles. The data presented in this study suggest that Vps33b is involved in α‐granule formation possibly by facilitating the Vwf‐positive vesicular trafficking to α‐granule‐related vacuoles in megakaryocytes. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

[1]  S. Smyth,et al.  Characterization of a Novel Integrin Binding Protein, VPS33B, Which Is Important for Platelet Activation and In Vivo Thrombosis and Hemostasis , 2015, Circulation.

[2]  P. van der Sluijs,et al.  Vps33B is required for delivery of endocytosed cargo to lysosomes , 2015, Traffic.

[3]  Rik van der Kant,et al.  Characterization of the Mammalian CORVET and HOPS Complexes and Their Modular Restructuring for Endosome Specificity* , 2015, The Journal of Biological Chemistry.

[4]  F. Rieux-Laucat,et al.  VPS33B regulates protein sorting into and maturation of α-granule progenitor organelles in mouse megakaryocytes. , 2015, Blood.

[5]  Junfeng Zhang,et al.  Arthrogryposis–renal dysfunction–cholestasis (ARC) syndrome: from molecular genetics to clinical features , 2014, Italian Journal of Pediatrics.

[6]  A. Fischer,et al.  Megakaryocyte-specific Profilin1-deficiency alters microtubule stability and causes a Wiskott–Aldrich syndrome-like platelet defect , 2014, Nature Communications.

[7]  X. Darzacq,et al.  The SNARE Sec22b has a non-fusogenic function in plasma membrane expansion , 2014, Nature Cell Biology.

[8]  V. Faundez,et al.  Vps33b pathogenic mutations preferentially affect VIPAS39/SPE-39-positive endosomes. , 2013, Human molecular genetics.

[9]  C. Hawkins,et al.  Abnormal megakaryocyte development and platelet function in Nbeal2(-/-) mice. , 2013, Blood.

[10]  P. Nurden,et al.  Gray platelet syndrome and defective thrombo-inflammation in Nbeal2-deficient mice. , 2013, The Journal of clinical investigation.

[11]  D. Owen,et al.  Structural basis of Vps33A recruitment to the human HOPS complex by Vps16 , 2013, Proceedings of the National Academy of Sciences.

[12]  P. Jeffrey,et al.  Crystal Structures of the Sec1/Munc18 (SM) Protein Vps33, Alone and Bound to the Homotypic Fusion and Vacuolar Protein Sorting (HOPS) Subunit Vps16* , 2013, PloS one.

[13]  J. Rizo,et al.  Reconstitution of the Vital Functions of Munc18 and Munc13 in Neurotransmitter Release , 2013, Science.

[14]  P. Nurden,et al.  The Nbeal2−/− mouse as a model for the gray platelet syndrome , 2013, Rare diseases.

[15]  J. Parkinson,et al.  The VPS33B-binding protein VPS16B is required in megakaryocyte and platelet α-granule biogenesis. , 2012, Blood.

[16]  T. Südhof,et al.  The membrane fusion enigma: SNAREs, Sec1/Munc18 proteins, and their accomplices--guilty as charged? , 2012, Annual review of cell and developmental biology.

[17]  J. Rizo.  Staging Membrane Fusion , 2012, Science.

[18]  J. Enninga,et al.  Sec22b Regulates Phagosomal Maturation and Antigen Crosspresentation by Dendritic Cells , 2011, Cell.

[19]  J. Parkinson,et al.  Mutations in NBEAL2, encoding a BEACH protein, cause gray platelet syndrome , 2011, Nature Genetics.

[20]  Gregory E. Jordan,et al.  Exome sequencing identifies NBEAL2 as the causative gene for Gray Platelet Syndrome , 2011, Nature Genetics.

[21]  J. Mullikin,et al.  NBEAL2 is mutated in Gray Platelet Syndrome and is required for biogenesis of platelet alpha-granules , 2011, Nature Genetics.

[22]  J. Freedman,et al.  Platelets and the immune continuum , 2011, Nature Reviews Immunology.

[23]  Laurie J. Gay,et al.  Contribution of platelets to tumour metastasis , 2011, Nature Reviews Cancer.

[24]  Guoqiang Chen,et al.  PTEN regulates collagen-induced platelet activation. , 2010, Blood.

[25]  H. Mandel,et al.  Mutations in VIPAR cause an arthrogryposis, renal dysfunction and cholestasis syndrome phenotype with defects in epithelial polarization , 2010, Nature Genetics.

[26]  E. Battinelli,et al.  Selective sorting of alpha‐granule proteins , 2009, Journal of thrombosis and haemostasis : JTH.

[27]  R. Flaumenhaft,et al.  Platelet alpha-granules: basic biology and clinical correlates. , 2009, Blood reviews.

[28]  E. Yu,et al.  Clinical Characteristics and VPS33B Mutations in Patients With ARC Syndrome , 2009, Journal of pediatric gastroenterology and nutrition.

[29]  R. Sougrat,et al.  Molecular investigations to improve diagnostic accuracy in patients with ARC syndrome , 2009, Human mutation.

[30]  B. Furie,et al.  Mechanisms of thrombus formation. , 2008, The New England journal of medicine.

[31]  D. Chitayat,et al.  Requirement of VPS33B, a member of the Sec1/Munc18 protein family, in megakaryocyte and platelet alpha-granule biogenesis. , 2005, Blood.

[32]  Joseph E Italiano,et al.  Mechanisms of organelle transport and capture along proplatelets during platelet production. , 2005, Blood.

[33]  J. Rohrer,et al.  Drosophila Vps16A is required for trafficking to lysosomes and biogenesis of pigment granules , 2005, Journal of Cell Science.

[34]  Mark A. Hall,et al.  In vivo fate-tracing studies using the Scl stem cell enhancer: embryonic hematopoietic stem cells significantly contribute to adult hematopoiesis. , 2004, Blood.

[35]  Richard C Trembath,et al.  Mutations in VPS33B, encoding a regulator of SNARE-dependent membrane fusion, cause arthrogryposis–renal dysfunction–cholestasis (ARC) syndrome , 2004, Nature Genetics.

[36]  D. Camerini,et al.  Platelet Chemokines and Chemokine Receptors: Linking Hemostasis, Inflammation, and Host Defense , 2003, Microcirculation.

[37]  R. Spritz,et al.  The mouse organellar biogenesis mutant buff results from a mutation in Vps33a, a homologue of yeast vps33 and Drosophila carnation , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[38]  A. Leavitt,et al.  Megakaryocytes derived from embryonic stem cells implicate CalDAG-GEFI in integrin signaling , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[39]  G. Reed,et al.  Molecular mechanisms of platelet exocytosis: insights into the "secrete" life of thrombocytes. , 2000, Blood.

[40]  E. Cramer,et al.  Megakaryocyte dense granule components are sorted in multivesicular bodies. , 2000, Blood.

[41]  N. Debili,et al.  Multivesicular Bodies Are an Intermediate Stage in the Formation of Platelet α-Granules , 1998 .

[42]  E. Cramer,et al.  Gray platelet syndrome. Dissociation between abnormal sorting in megakaryocyte alpha-granules and normal sorting in Weibel-Palade bodies of endothelial cells. , 1993, The Journal of clinical investigation.

[43]  J. Maclouf,et al.  Gray platelet syndrome: alpha-granule deficiency. Its influence on platelet function. , 1981, The Journal of laboratory and clinical medicine.

[44]  T. Reese.  Synaptic vesicle exocytosis. , 1981, JAMA.

[45]  S. Niewiarowski.  Proteins Secreted by the Platelet , 1977, Thrombosis and Haemostasis.

[46]  Junling Liu,et al.  The 3 subunit of the integrin IIb3 regulates IIb-mediated outside-in signaling , 2016 .

[47]  N. Debili,et al.  Multivesicular bodies are an intermediate stage in the formation of platelet alpha-granules. , 1998, Blood.

[48]  R. Scheller,et al.  Mammalian homologues of yeast vacuolar protein sorting (vps) genes implicated in Golgi-to-lysosome trafficking. , 1996, Gene.

[49]  T. Miki-Noumura,et al.  Extrusion of rotating microtubules on the dynein-track from a microtubule-dynein gamma-complex. , 1995, Cell motility and the cytoskeleton.

[50]  J. Slot,et al.  Immunocytochemical localization of platelet granule proteins. , 1989, Methods in enzymology.

[51]  J. Slot,et al.  [26] Immunocytochemical localization of platelet granule proteins , 1989 .

[52]  W. Vainchenker,et al.  Gray platelet syndrome: immunoelectron microscopic localization of fibrinogen and von Willebrand factor in platelets and megakaryocytes. , 1985, Blood.

[53]  C. Soria,et al.  Specific protein and glycoprotein deficiencies in platelets isolated from two patients with the gray platelet syndrome. , 1982, Blood.

[54]  W. Vainchenker,et al.  Defective alpha-granule production in megakaryocytes from gray platelet syndrome: ultrastructural studies of bone marrow cells and megakaryocytes growing in culture from blood precursors. , 1981, The American journal of pathology.