A positive signal prevents secretory membrane cargo from recycling between the Golgi and the ER

The Golgi complex and ER are dynamically connected by anterograde and retrograde trafficking pathways. To what extent and by what mechanism outward‐bound cargo proteins escape retrograde trafficking has been poorly investigated. Here, we analysed the behaviour of several membrane proteins at the ER/Golgi interface in live cells. When Golgi‐to‐plasma membrane transport was blocked, vesicular stomatitis virus glycoprotein (VSVG), which bears an ER export signal, accumulated in the Golgi, whereas an export signal‐deleted version of VSVG attained a steady state determined by the balance of retrograde and anterograde traffic. A similar behaviour was displayed by EGF receptor and by a model tail‐anchored protein, whose retrograde traffic was slowed by addition of VSVG's export signal. Retrograde trafficking was energy‐ and Rab6‐dependent, and Rab6 inhibition accelerated signal‐deleted VSVG's transport to the cell surface. Our results extend the dynamic bi‐directional relationship between the Golgi and ER to include surface‐directed proteins, uncover an unanticipated role for export signals at the Golgi complex, and identify recycling as a novel factor that regulates cargo transport out of the early secretory pathway.

[1]  S. Colombo,et al.  Nicotine-Modulated Subunit Stoichiometry Affects Stability and Trafficking of α3β4 Nicotinic Receptor , 2013, The Journal of Neuroscience.

[2]  A. Fornili,et al.  A pH-Regulated Quality Control Cycle for Surveillance of Secretory Protein Assembly , 2013, Molecular cell.

[3]  A. Spang Retrograde traffic from the Golgi to the endoplasmic reticulum. , 2013, Cold Spring Harbor perspectives in biology.

[4]  Kota Sato,et al.  A New Class of Endoplasmic Reticulum Export Signal ΦXΦXΦ for Transmembrane Proteins and Its Selective Interaction with Sec24C* , 2013, The Journal of Biological Chemistry.

[5]  A. Luini,et al.  The KDEL receptor couples to Gαq/11 to activate Src kinases and regulate transport through the Golgi , 2012, The EMBO journal.

[6]  Jeffrey A Kamykowski,et al.  Electron Tomography Reveals Rab6 Is Essential to the Trafficking of trans‐Golgi Clathrin and COPI‐Coated Vesicles and the Maintenance of Golgi Cisternal Number , 2012, Traffic.

[7]  Franck Perez,et al.  Synchronization of secretory protein traffic in populations of cells , 2012, Nature Methods.

[8]  W. Balch,et al.  Di-acidic Motifs in the Membrane-distal C Termini Modulate the Transport of Angiotensin II Receptors from the Endoplasmic Reticulum to the Cell Surface* , 2011, The Journal of Biological Chemistry.

[9]  G. Beznoussenko,et al.  GRASP65 and GRASP55 Sequentially Promote the Transport of C-terminal Valine-bearing Cargos to and through the Golgi Complex* , 2009, The Journal of Biological Chemistry.

[10]  K. Hirschberg,et al.  The length of cargo-protein transmembrane segments drives secretory transport by facilitating cargo concentration in export domains , 2009, Journal of Cell Science.

[11]  M. Robitaille,et al.  A Single Conserved Leucine Residue on the First Intracellular Loop Regulates ER Export of G Protein‐Coupled Receptors , 2009, Traffic.

[12]  J. Mancias,et al.  Structural basis of cargo membrane protein discrimination by the human COPII coat machinery , 2008, The EMBO journal.

[13]  Robert D. Phair,et al.  Transport through the Golgi Apparatus by Rapid Partitioning within a Two-Phase Membrane System , 2008, Cell.

[14]  M. Francolini,et al.  Transmembrane domain–dependent partitioning of membrane proteins within the endoplasmic reticulum , 2008, The Journal of cell biology.

[15]  J. Christopher Fromme,et al.  The genetic basis of a craniofacial disease provides insight into COPII coat assembly. , 2007, Developmental cell.

[16]  D. Spasic,et al.  Rer1p competes with APH-1 for binding to nicastrin and regulates γ-secretase complex assembly in the early secretory pathway , 2007, The Journal of cell biology.

[17]  D. Spasic,et al.  Rer1p Competes with Aph-1 for Binding to Nicastrin and Regulates gamma-secretase Complex Assembly , 2007 .

[18]  J. Riordan,et al.  COPII-dependent export of cystic fibrosis transmembrane conductance regulator from the ER uses a di-acidic exit code , 2004, The Journal of cell biology.

[19]  H. Sitte,et al.  Two Discontinuous Segments in the Carboxyl Terminus Are Required for Membrane Targeting of the Rat γ-Aminobutyric Acid Transporter-1 (GAT1)* , 2004, Journal of Biological Chemistry.

[20]  K. Hahn,et al.  The δ subunit of AP-3 is required for efficient transport of VSV-G from the trans-Golgi network to the cell surface , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[21]  E. Pedrazzini,et al.  Trafficking of tail-anchored proteins: transport from the endoplasmic reticulum to the plasma membrane and sorting between surface domains in polarised epithelial cells. , 2002, Journal of cell science.

[22]  A. Luini,et al.  Small cargo proteins and large aggregates can traverse the Golgi by a common mechanism without leaving the lumen of cisternae , 2001, The Journal of cell biology.

[23]  Takashi Saito,et al.  The KDEL receptor mediates a retrieval mechanism that contributes to quality control at the endoplasmic reticulum , 2001, The EMBO journal.

[24]  Y. Jan,et al.  Role of ER export signals in controlling surface potassium channel numbers. , 2001, Science.

[25]  J. Riordan,et al.  Traffic Pattern of Cystic Fibrosis Transmembrane Regulator through the Early Exocytic Pathway , 2000, Traffic.

[26]  A. Linstedt,et al.  Potential role for protein kinases in regulation of bidirectional endoplasmic reticulum-to-Golgi transport revealed by protein kinase inhibitor H89. , 2000, Molecular biology of the cell.

[27]  C. Sevier,et al.  Efficient export of the vesicular stomatitis virus G protein from the endoplasmic reticulum requires a signal in the cytoplasmic tail that includes both tyrosine-based and di-acidic motifs. , 2000, Molecular biology of the cell.

[28]  Ludger Johannes,et al.  Rab6 Coordinates a Novel Golgi to ER Retrograde Transport Pathway in Live Cells , 1999, The Journal of cell biology.

[29]  R. Pepperkok,et al.  Evidence for a COP-I-independent transport route from the Golgi complex to the endoplasmic reticulum , 1999, Nature Cell Biology.

[30]  W. Balch,et al.  A Di-acidic (DXE) Code Directs Concentration of Cargo during Export from the Endoplasmic Reticulum* , 1999, The Journal of Biological Chemistry.

[31]  J. Rothman,et al.  Coupling of Coat Assembly and Vesicle Budding to Packaging of Putative Cargo Receptors , 1999, Cell.

[32]  A. Sorkin,et al.  Endocytosis of Functional Epidermal Growth Factor Receptor-Green Fluorescent Protein Chimera* , 1998, The Journal of Biological Chemistry.

[33]  E. Stelzer,et al.  Recycling of Golgi-resident Glycosyltransferases through the ER Reveals a Novel Pathway and Provides an Explanation for Nocodazole-induced Golgi Scattering , 1998, The Journal of cell biology.

[34]  P. Cosson,et al.  New COP1‐binding motifs involved in ER retrieval , 1998, The EMBO journal.

[35]  J. Lippincott-Schwartz,et al.  Retrograde Transport of Golgi-localized Proteins to the ER , 1998, The Journal of cell biology.

[36]  E. Berger,et al.  Localization of three human polypeptide GalNAc-transferases in HeLa cells suggests initiation of O-linked glycosylation throughout the Golgi apparatus. , 1998, Journal of cell science.

[37]  L. Johannes,et al.  Retrograde Transport of KDEL-bearing B-fragment of Shiga Toxin* , 1997, The Journal of Biological Chemistry.

[38]  W. Balch,et al.  A di-acidic signal required for selective export from the endoplasmic reticulum. , 1997, Science.

[39]  E. Pedrazzini,et al.  A mutant cytochrome b5 with a lengthened membrane anchor escapes from the endoplasmic reticulum and reaches the plasma membrane. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[40]  L. Swift Assembly of very low density lipoproteins in rat liver: a study of nascent particles recovered from the rough endoplasmic reticulum. , 1995, Journal of lipid research.

[41]  M. Roth,et al.  The basolateral targeting signal in the cytoplasmic domain of glycoprotein G from vesicular stomatitis virus resembles a variety of intracellular targeting motifs related by primary sequence but having diverse targeting activities. , 1994, The Journal of biological chemistry.

[42]  R Pepperkok,et al.  The intracellular mobility of a viral membrane glycoprotein measured by confocal microscope fluorescence recovery after photobleaching. , 1994, Journal of cell science.

[43]  P. Cosson,et al.  Coatomer interaction with di-lysine endoplasmic reticulum retention motifs. , 1994, Science.

[44]  J. Rothman,et al.  Hydrolysis of bound GTP by ARF protein triggers uncoating of Golgi- derived COP-coated vesicles , 1993, The Journal of cell biology.

[45]  M. Roth,et al.  Vesicular stomatitis virus glycoprotein contains a dominant cytoplasmic basolateral sorting signal critically dependent upon a tyrosine. , 1993, The Journal of biological chemistry.

[46]  R. Doms,et al.  Brefeldin A redistributes resident and itinerant Golgi proteins to the endoplasmic reticulum , 1989, The Journal of cell biology.

[47]  J. Lippincott-Schwartz,et al.  Rapid redistribution of Golgi proteins into the ER in cells treated with brefeldin A: Evidence for membrane cycling from Golgi to ER , 1989, Cell.

[48]  H. Hauri,et al.  Identification, by a monoclonal antibody, of a 53-kD protein associated with a tubulo-vesicular compartment at the cis-side of the Golgi apparatus , 1988, The Journal of cell biology.

[49]  M. Lane,et al.  Assembly of very low density lipoprotein in the hepatocyte. Differential transport of apoproteins through the secretory pathway. , 1988, The Journal of biological chemistry.

[50]  K. Olden,et al.  Variability in transport rates of secretory glycoproteins through the endoplasmic reticulum and Golgi in human hepatoma cells. , 1985, The Journal of biological chemistry.

[51]  J. Rose,et al.  A single amino acid substitution in a hydrophobic domain causes temperature-sensitive cell-surface transport of a mutant viral glycoprotein , 1985, Journal of virology.

[52]  K. Simons,et al.  Reduced temperature prevents transfer of a membrane glycoprotein to the cell surface but does not prevent terminal glycosylation , 1983, Cell.