Sec7p directs the transitions required for yeast Golgi biogenesis.
Endoplasmic reticulum (ER)-to-Golgi traffic in yeast proceeds by the maturation of membrane compartments from post-ER vesicles to intermediate small vesicle tubular clusters (VTCs) to Golgi nodular membrane networks (Morin-Ganet et al., Traffic 2000; 1: 56-68). The balance between ER and Golgi compartments is maintained by COPII- and COPI-mediated anterograde and retrograde traffic, which are dependent on Sec7p and ARF function. The sec7-4 temperature-sensitive allele is a mutation in the highly conserved Sec7 domain (Sec7d) found in all ARF-guanine nucleotide exchange factor proteins. Post-ER trafficking is rapidly inactivated in sec7-4 mutant yeast at the restrictive temperature. This conditional defect prevented the normal production of VTCs and instead generated Golgi-like tubes emanating from the ER exit sites. These tubes progressively developed into stacked cisternae defining the landmark sec7 mutant phenotype. Consistent with the in vivo results, a Sec7d peptide inhibited ER-to-Golgi transport and displaced Sec7p from its membrane anchor in vitro. The similarities in the consequences of inactivating Sec7p or ARFs in vivo was revealed by genetic disruption of yeast ARFs or by addition of brefeldin A (BFA) to whole cells. These treatments, as in sec7-4 yeast, affected the morphology of membrane compartments in the ER-Golgi transition. Further evidence for Sec7p involvement in the transition for Golgi biogenesis was revealed by in vitro binding between distinct domains of Sec7p with ARFs, COPI and COPII coat proteins. These results suggest that Sec7p coordinates membrane transitions in Golgi biogenesis by directing and scaffolding the binding and disassembly of coat protein complexes to membranes, both at the VTC transition from ER exit sites to form Golgi elements and for later events in Golgi maturation.
Deitz
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Program in Molecular Biology and CU Cancer Center, Department of Cellular and Structural Biology, Box B-111, University of Colorado Health Sciences Center, Denver, CO 80262, USA.
Rambourg
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Képès
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Franzusoff
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D000483:Alleles; D000595:Amino Acid Sequence; D020126:Brefeldin A / Q000494:pharmacology; D022181:COP-Coated Vesicles / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D002474:Cell-Free System; D003001:Cloning, Molecular; D020755:Coat Protein Complex I / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D005838:Genotype; D005982:Glutathione Transferase / Q000378:metabolism; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D020662:Guanine Nucleotide Exchange Factors; D007700:Kinetics; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D010641:Phenotype; D011233:Precipitin Tests; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000502:physiology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D013997:Time Factors
D020755:Coat Protein Complex I; D005656:Fungal Proteins; D020662:Guanine Nucleotide Exchange Factors; D010455:Peptides; D011500:Protein Synthesis Inhibitors; D011993:Recombinant Fusion Proteins; C066755:Sec7 guanine nucleotide exchange factors; D020126:Brefeldin A; D005982:Glutathione Transferase