Home LiteratureArticle Details
PMID: 7876304 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Human SEC13Rp functions in yeast and is located on transport vesicles budding from the endoplasmic reticulum.

The Journal of cell biology ·Vol. 128 ·No. 5 ·1995-03-00 ·Pages 769-77

Shaywitz DA, Orci L, Ravazzola M, Swaroop A, Kaiser CA

Abstract

In the yeast Saccharomyces cerevisiae, Sec13p is required for intracellular protein transport from the ER to the Golgi apparatus, and has also been identified as a component of the COPII vesicle coat structure. Recently, a human cDNA encoding a protein 53% identical to yeast Sec13p has been isolated. In this report, we apply the genetic assays of complementation and synthetic lethality to demonstrate the conservation of function between this human protein, designated SEC13Rp, and yeast Sec13p. We show that two reciprocal human/yeast fusion constructs, encoding the NH2-terminal half of one protein and the COOH-terminal half of the other, can each complement the secretion defect of a sec13-1 mutant at 36 degrees C. The chimera encoding the NH2-terminal half of the yeast protein and the COOH-terminal half of the human protein is also able to complement a SEC13 deletion. Overexpression of either the entire human SEC13Rp protein or the chimera encoding the NH2-terminal half of the human protein and the COOH-terminal half of the yeast protein inhibits the growth of a sec13-1 mutant at 24 degrees C; this growth inhibition is not seen in a wild-type strain nor in other sec mutants, suggesting that the NH2-terminal half of SEC13Rp may compete with Sec13-1p for a common target. We show by immunoelectronmicroscopy of mammalian cells that SEC13Rp (like the putative mammalian homologues of the COPII subunits Sar1p and Sec23p) resides in the region of the transitional ER. We also show that the distribution of SEC13Rp is not affected by brefeldin A treatment. This report presents the first demonstration of a putative mammalian COPII component functioning in yeast, and highlights a potentially useful approach for the study of conserved mammalian proteins in a genetically tractable system.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Biological Transport CHO Cells Cell Compartmentation Cells, Cultured Cricetinae Endoplasmic Reticulum/physiology Fluorescent Antibody Technique Fungal Proteins/genetics,isolation & purification,physiology Genetic Complementation Test Humans Intracellular Membranes/physiology Membrane Proteins/genetics,isolation & purification,physiology Microscopy, Immunoelectron Molecular Sequence Data Mutation Nuclear Pore Complex Proteins Pancreas/ultrastructure Rats Recombinant Fusion Proteins Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins
Chemicals
Fungal Proteins Membrane Proteins Nuclear Pore Complex Proteins Recombinant Fusion Proteins SEC13 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Shaywitz D A
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Orci L
Ravazzola M
Swaroop A
Kaiser C A
References (49)
49 references, click to expand
  1. Mammalian Sec23p homologue is restricted to the endoplasmic reticulum transitional cytoplasm.
    Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8611-5 PMID: 1924322
  2. The yeast SEC17 gene product is functionally equivalent to mammalian alpha-SNAP protein.
    J Biol Chem. 1992 Jun 15;267(17):12106-15 PMID: 1601878
  3. Inhibition by brefeldin A of a Golgi membrane enzyme that catalyses exchange of guanine nucleotide bound to ARF.
    Nature. 1992 Nov 26;360(6402):352-4 PMID: 1448152
  4. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  5. Binding of coatomer to Golgi membranes requires ADP-ribosylation factor.
    J Biol Chem. 1993 Jun 5;268(16):12083-9 PMID: 8505331
  6. "BFA bodies": a subcompartment of the endoplasmic reticulum.
    Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11089-93 PMID: 8248213
  7. SEC21 is a gene required for ER to Golgi protein transport that encodes a subunit of a yeast coatomer.
    Nature. 1992 Dec 10;360(6404):603-5 PMID: 1461285
  8. Fusions to staphylococcal protein A.
    Methods Enzymol. 1990;185:144-61 PMID: 2199777
  9. Construction of a new family of high efficiency bacterial expression vectors: identification of cDNA clones coding for human liver proteins.
    EMBO J. 1984 Jun;3(6):1429-34 PMID: 6086324
  10. Distinct sets of SEC genes govern transport vesicle formation and fusion early in the secretory pathway.
    Cell. 1990 May 18;61(4):723-33 PMID: 2188733
  11. Dominant inhibitory mutants of ARF1 block endoplasmic reticulum to Golgi transport and trigger disassembly of the Golgi apparatus.
    J Biol Chem. 1994 Jan 14;269(2):1437-48 PMID: 8288610
  12. Molecular dissection of the secretory pathway.
    Nature. 1992 Jan 30;355(6359):409-15 PMID: 1734280
  13. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Methods Enzymol. 1987;154:367-82 PMID: 3323813
  14. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  15. Cloning of a yeast U1 snRNP 70K protein homologue: functional conservation of an RNA-binding domain between humans and yeast.
    EMBO J. 1991 Sep;10(9):2627-34 PMID: 1714384
  16. Early stages in the yeast secretory pathway are required for transport of carboxypeptidase Y to the vacuole.
    Cell. 1982 Sep;30(2):439-48 PMID: 6754086
  17. Identification of revertants for the cystic fibrosis delta F508 mutation using STE6-CFTR chimeras in yeast.
    Cell. 1993 Apr 23;73(2):335-46 PMID: 7682896
  18. A Highly Specific Complementary Lethal System in Drosophila Melanogaster.
    Genetics. 1956 Jan;41(1):118-23 PMID: 17247604
  19. Ultrastructural localization of intracellular antigens by the use of protein A-gold complex.
    J Histochem Cytochem. 1978 Dec;26(12):1074-81 PMID: 366014
  20. Beta-COP is essential for biosynthetic membrane transport from the endoplasmic reticulum to the Golgi complex in vivo.
    Cell. 1993 Jul 16;74(1):71-82 PMID: 8334707
  21. Identification of a mammalian gene structurally and functionally related to the CDC25 gene of Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7100-4 PMID: 1379731
  22. Immunochemistry on ultrathin frozen sections.
    Histochem J. 1980 Jul;12(4):381-403 PMID: 7440248
  23. A novel GTP-binding protein, Sar1p, is involved in transport from the endoplasmic reticulum to the Golgi apparatus.
    J Cell Biol. 1989 Dec;109(6 Pt 1):2677-91 PMID: 2512296
  24. Vesicle-mediated protein sorting.
    Annu Rev Biochem. 1992;61:471-516 PMID: 1497318
  25. Stepwise assembly of functionally active transport vesicles.
    Cell. 1993 Dec 3;75(5):1015-25 PMID: 8252615
  26. ADP-ribosylation factor, a small GTP-binding protein, is required for binding of the coatomer protein beta-COP to Golgi membranes.
    Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6408-12 PMID: 1631136
  27. Genes conserved in yeast and humans.
    Hum Mol Genet. 1994;3 Spec No:1509-17 PMID: 7849746
  28. Sar1 promotes vesicle budding from the endoplasmic reticulum but not Golgi compartments.
    J Cell Biol. 1994 Apr;125(1):51-65 PMID: 8138575
  29. En bloc incorporation of coatomer subunits during the assembly of COP-coated vesicles.
    J Cell Biol. 1994 Mar;124(6):883-92 PMID: 8132710
  30. Molecular characterization of a novel human gene, SEC13R, related to the yeast secretory pathway gene SEC13, and mapping to a conserved linkage group on human chromosome 3p24-p25 and mouse chromosome 6.
    Hum Mol Genet. 1994 Aug;3(8):1281-6 PMID: 7987303
  31. Coated vesicle assembly in the Golgi requires only coatomer and ARF proteins from the cytosol.
    Nature. 1993 Aug 19;364(6439):732-4 PMID: 8355790
  32. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains.
    Genetics. 1987 Aug;116(4):541-5 PMID: 3305158
  33. The fine structure of the gene.
    Sci Am. 1962 Jan;206:70-84 PMID: 13867419
  34. Identification and differential expression of yeast SEC23-related gene (Msec23) in mouse tissues.
    FEBS Lett. 1993 Jan 4;315(2):193-6 PMID: 8417978
  35. ADP-ribosylation factor and coatomer couple fusion to vesicle budding.
    J Cell Biol. 1994 Feb;124(4):415-24 PMID: 8106543
  36. Budding from Golgi membranes requires the coatomer complex of non-clathrin coat proteins.
    Nature. 1993 Apr 15;362(6421):648-52 PMID: 8464517
  37. Two distinct populations of ARF bound to Golgi membranes.
    J Cell Biol. 1993 May;121(4):751-60 PMID: 8491770
  38. The Sec13p complex and reconstitution of vesicle budding from the ER with purified cytosolic proteins.
    EMBO J. 1993 Nov;12(11):4073-82 PMID: 8223424
  39. Cytosolic Sec13p complex is required for vesicle formation from the endoplasmic reticulum in vitro.
    J Cell Biol. 1993 Feb;120(4):865-75 PMID: 8432727
  40. Genetic analysis of the mitotic transmission of minichromosomes.
    Cell. 1985 Feb;40(2):393-403 PMID: 3881185
  41. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance.
    Mol Gen Genet. 1984;197(2):345-6 PMID: 6394957
  42. Identification of 23 complementation groups required for post-translational events in the yeast secretory pathway.
    Cell. 1980 Aug;21(1):205-15 PMID: 6996832
  43. COPII: a membrane coat formed by Sec proteins that drive vesicle budding from the endoplasmic reticulum.
    Cell. 1994 Jun 17;77(6):895-907 PMID: 8004676
  44. Hydrolysis of bound GTP by ARF protein triggers uncoating of Golgi-derived COP-coated vesicles.
    J Cell Biol. 1993 Dec;123(6 Pt 1):1365-71 PMID: 8253837
  45. Beta-COP is essential for transport of protein from the endoplasmic reticulum to the Golgi in vitro.
    J Cell Biol. 1993 Sep;122(6):1155-67 PMID: 8376457
  46. Intracellular aspects of the process of protein synthesis.
    Science. 1975 Aug 1;189(4200):347-58 PMID: 1096303
  47. Sec23p and a novel 105-kDa protein function as a multimeric complex to promote vesicle budding and protein transport from the endoplasmic reticulum.
    Mol Biol Cell. 1992 Jun;3(6):667-76 PMID: 1498369
  48. Genetic analysis of the yeast cytoskeleton.
    Annu Rev Genet. 1987;21:259-84 PMID: 3327466
  49. BRG1 contains a conserved domain of the SWI2/SNF2 family necessary for normal mitotic growth and transcription.
    Nature. 1993 Nov 11;366(6451):170-4 PMID: 8232556
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
Published
1995-03-00
Pages
769-77
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120388
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com