The DHR1 domain of DOCK180 binds to SNX5 and regulates cation-independent mannose 6-phosphate receptor transport.

DOCK180 is the archetype of the DOCK180-family guanine nucleotide exchange factor for small GTPases Rac1 and Cdc42. DOCK180-family proteins share two conserved domains, called DOCK homology region (DHR)-1 and -2. Although the function of DHR2 is to activate Rac1, DHR1 is required for binding to phosphoinositides. To better understand the function of DHR1, we searched for its binding partners by direct nanoflow liquid chromatography/tandem mass spectrometry, and we identified sorting nexins (SNX) 1, 2, 5, and 6, which make up a multimeric protein complex mediating endosome-to-trans-Golgi-network (TGN) retrograde transport of the cation-independent mannose 6-phosphate receptor (CI-MPR). Among these SNX proteins, SNX5 was coimmunoprecipitated with DOCK180 most efficiently. In agreement with this observation, DOCK180 colocalized with SNX5 at endosomes. The RNA interference-mediated knockdowns of SNX5 and DOCK180, but not Rac1, resulted in the redistribution of CI-MPR from TGN to endosomes. Furthermore, expression of the DOCK180 DHR1 domain was sufficient to restore the perturbed CI-MPR distribution in DOCK180 knockdown cells. These data suggest that DOCK180 regulates CI-MPR trafficking via SNX5 and that this function is independent of its guanine nucleotide exchange factor activity toward Rac1.

[1]  J. Kawai,et al.  Mutation of Dock5, a member of the guanine exchange factor Dock180 superfamily, in the rupture of lens cataract mouse. , 2008, Experimental eye research.

[2]  S. Pfeffer,et al.  A syntaxin 10–SNARE complex distinguishes two distinct transport routes from endosomes to the trans-Golgi in human cells , 2008, The Journal of cell biology.

[3]  L. Johannes,et al.  The retromer component sorting nexin-1 is required for efficient retrograde transport of Shiga toxin from early endosome to the trans Golgi network , 2007, Journal of Cell Science.

[4]  T. Meyer,et al.  An essential role for the SHIP2-dependent negative feedback loop in neuritogenesis of nerve growth factor–stimulated PC12 cells , 2007, The Journal of cell biology.

[5]  T. Wassmer,et al.  A loss-of-function screen reveals SNX5 and SNX6 as potential components of the mammalian retromer , 2006, Journal of Cell Science.

[6]  Pietro De Camilli,et al.  Phosphoinositides in cell regulation and membrane dynamics , 2006, Nature.

[7]  R. Teasdale,et al.  Visualisation of macropinosome maturation by the recruitment of sorting nexins , 2006, Journal of Cell Science.

[8]  Mitsuko Watabe-Uchida,et al.  The Rac Activator DOCK7 Regulates Neuronal Polarity through Local Phosphorylation of Stathmin/Op18 , 2006, Neuron.

[9]  M. Matsuda,et al.  The amino‐terminal B‐Raf‐specific region mediates calcium‐dependent homo‐ and hetero‐dimerization of Raf , 2006, The EMBO journal.

[10]  Stephen Magill,et al.  Inhibitory regulation of EGF receptor degradation by sorting nexin 5. , 2006, Biochemical and biophysical research communications.

[11]  E. Nishida,et al.  Dynamics of the Ras/ERK MAPK Cascade as Monitored by Fluorescent Probes* , 2006, Journal of Biological Chemistry.

[12]  Bianca Habermann,et al.  Dynamin and the actin cytoskeleton cooperatively regulate plasma membrane invagination by BAR and F-BAR proteins. , 2005, Developmental cell.

[13]  R. Teasdale,et al.  A Novel Mammalian Retromer Component, Vps26B , 2005, Traffic.

[14]  B. Peter,et al.  Sorting nexin-2 is associated with tubular elements of the early endosome, but is not essential for retromer-mediated endosome-to-TGN transport , 2005, Journal of Cell Science.

[15]  K. Vuori,et al.  A novel and evolutionarily conserved PtdIns(3,4,5)P3-binding domain is necessary for DOCK180 signalling , 2005, Nature Cell Biology.

[16]  M. Seaman,et al.  Recycle your receptors with retromer. , 2005, Trends in cell biology.

[17]  R. Teasdale,et al.  Sorting nexin 5 is localized to a subdomain of the early endosomes and is recruited to the plasma membrane following EGF stimulation , 2004, Journal of Cell Science.

[18]  Judith Klumperman,et al.  Sorting Nexin-1 Mediates Tubular Endosome-to-TGN Transport through Coincidence Sensing of High- Curvature Membranes and 3-Phosphoinositides , 2004, Current Biology.

[19]  J. Trejo,et al.  A role for sorting nexin 2 in epidermal growth factor receptor down-regulation: evidence for distinct functions of sorting nexin 1 and 2 in protein trafficking. , 2004, Molecular biology of the cell.

[20]  M. Seaman.  Cargo-selective endosomal sorting for retrieval to the Golgi requires retromer , 2004, The Journal of cell biology.

[21]  Bianca Habermann,et al.  The BAR‐domain family of proteins: a case of bending and binding? , 2004, EMBO reports.

[22]  B. Peter,et al.  BAR Domains as Sensors of Membrane Curvature: The Amphiphysin BAR Structure , 2004, Science.

[23]  H. Hanafusa,et al.  Refractory nature of normal human diploid fibroblasts with respect to oncogene-mediated transformation , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Hironori Katoh,et al.  RhoG activates Rac1 by direct interaction with the Dock180-binding protein Elmo , 2003, Nature.

[25]  D. Haber,et al.  DOCK4, a GTPase Activator, Is Disrupted during Tumorigenesis , 2003, Cell.

[26]  Jack E. Dixon,et al.  Sorting out the cellular functions of sorting nexins , 2003, Nature Reviews Molecular Cell Biology.

[27]  K. Vuori,et al.  Identification of an evolutionarily conserved superfamily of DOCK180-related proteins with guanine nucleotide exchange activity , 2002, Journal of Cell Science.

[28]  R. Teasdale,et al.  The Phox Homology (PX) Domain-dependent, 3-Phosphoinositide-mediated Association of Sorting Nexin-1 with an Early Sorting Endosomal Compartment Is Required for Its Ability to Regulate Epidermal Growth Factor Receptor Degradation* , 2002, The Journal of Biological Chemistry.

[29]  M. Schwartz,et al.  Zizimin1, a novel Cdc42 activator, reveals a new GEF domain for Rho proteins , 2002, Nature Cell Biology.

[30]  Tohru Natsume,et al.  A direct nanoflow liquid chromatography-tandem mass spectrometry system for interaction proteomics. , 2002, Analytical chemistry.

[31]  M. Hengartner,et al.  CED-12/ELMO, a Novel Member of the CrkII/Dock180/Rac Pathway, Is Required for Phagocytosis and Cell Migration , 2001, Cell.

[32]  Toshikazu Shirai,et al.  Haematopoietic cell-specific CDM family protein DOCK2 is essential for lymphocyte migration , 2001, Nature.

[33]  L. Karlsson,et al.  A large family of endosome-localized proteins related to sorting nexin 1. , 2001, The Biochemical journal.

[34]  A. Roberts,et al.  Sorting Nexin 6, a Novel SNX, Interacts with the Transforming Growth Factor-β Family of Receptor Serine-Threonine Kinases* , 2001, The Journal of Biological Chemistry.

[35]  S. Smerdon,et al.  The structural basis of Arfaptin-mediated cross-talk between Rac and Arf signalling pathways , 2001, Nature.

[36]  M. Matsuda,et al.  Membrane recruitment of DOCK180 by binding to PtdIns(3,4,5)P3. , 2001, The Biochemical journal.

[37]  M. Matsuda,et al.  Non-adherent cell-specific expression of DOCK2, a member of the human CDM-family proteins. , 1999, Biochimica et biophysica acta.

[38]  M. Matsuda,et al.  Activation of Rac1 by a Crk SH3-binding protein, DOCK180. , 1998, Genes & development.

[39]  Michiyuki Matsuda,et al.  Evidence That DOCK180 Up-regulates Signals from the CrkII-p130Cas Complex* , 1998, The Journal of Biological Chemistry.

[40]  Scott D. Emr,et al.  A Membrane Coat Complex Essential for Endosome-to-Golgi Retrograde Transport in Yeast , 1998, The Journal of cell biology.

[41]  H. Horvitz,et al.  C. elegans phagocytosis and cell-migration protein CED-5 is similar to human DOCK180 , 1998, Nature.

[42]  S. M. Abmayr,et al.  Drosophila myoblast city Encodes a Conserved Protein That Is Essential for Myoblast Fusion, Dorsal Closure, and Cytoskeletal Organization , 1997, The Journal of cell biology.

[43]  S. Emr,et al.  A sorting nexin-1 homologue, Vps5p, forms a complex with Vps17p and is required for recycling the vacuolar protein-sorting receptor. , 1997, Molecular biology of the cell.

[44]  S. Schmid,et al.  Regulation of receptor-mediated endocytosis by Rho and Rac , 1996, Nature.

[45]  R. Kurten,et al.  Enhanced Degradation of EGF Receptors by a Sorting Nexin, SNX1 , 1996, Science.

[46]  M. Shibuya,et al.  DOCK180, a major CRK-binding protein, alters cell morphology upon translocation to the cell membrane , 1996, Molecular and cellular biology.

[47]  Mitsuko Watabe-Uchida,et al.  Supplemental Experimental Procedures , 2022 .

[48]  I. Mellman,et al.  The biogenesis of lysosomes. , 1989, Annual review of cell biology.