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PMID: 6094591 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Mutations of the Rous sarcoma virus env gene that affect the transport and subcellular location of the glycoprotein products.

The Journal of cell biology ·Vol. 99 ·No. 6 ·1984-12-00 ·Pages 2011-23

Wills JW, Srinivas RV, Hunter E

Abstract

The envelope glycoproteins of Rous sarcoma virus (RSV), gp85 and gp37, are anchored in the membrane by a 27-amino acid, hydrophobic domain that lies adjacent to a 22-amino acid, cytoplasmic domain at the carboxy terminus of gp37. We have altered these cytoplasmic and transmembrane domains by introducing deletion mutations into the molecularly cloned sequences of a proviral env gene. The effects of the mutations on the transport and subcellular localization of the Rous sarcoma virus glycoproteins were examined in monkey (CV-1) cells using an SV40 expression vector. We found, on the one hand, that replacement of the nonconserved region of the cytoplasmic domain with a longer, unrelated sequence of amino acids (mutant C1) did not alter the rate of transport to the Golgi apparatus nor the appearance of the glycoprotein on the cell surface. Larger deletions, extending into the conserved region of the cytoplasmic domain (mutant C2), resulted in a slower rate of transport to the Golgi apparatus, but did not prevent transport to the cell surface. On the other hand, removal of the entire cytoplasmic and transmembrane domains (mutant C3) did block transport and therefore did not result in secretion of the truncated protein. Our results demonstrate that the C3 polypeptide was not transported to the Golgi apparatus, although it apparently remained in a soluble, nonanchored form in the lumen of the rough endoplasmic reticulum; therefore, it appears that this mutant protein lacks a functional sorting signal. Surprisingly, subcellular localization by internal immunofluorescence revealed that the C3 protein (unlike the wild type) did not accumulate on the nuclear membrane but rather in vesicles distributed throughout the cytoplasm. This observation suggests that the wild-type glycoproteins (and perhaps other membrane-bound or secreted proteins) are specifically transported to the nuclear membrane after their biosynthesis elsewhere in the rough endoplasmic reticulum.

MeSH Terms
Amino Acid Sequence Animals Avian Sarcoma Viruses/genetics Base Sequence Cell Line Chlorocebus aethiops DNA Restriction Enzymes Escherichia coli/genetics Genes Genes, Viral Glycoproteins/analysis,genetics Kidney Mutation Plasmids Subcellular Fractions/metabolism Viral Envelope Proteins/analysis,genetics
Chemicals
Glycoproteins Viral Envelope Proteins DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Wills J W
Srinivas R V
Hunter E
References (67)
67 references, click to expand
  1. Maturation of viral proteins in cells infected with temperature-sensitive mutants of vesicular stomatitis virus.
    J Virol. 1977 Mar;21(3):1149-58 PMID: 191642
  2. Studies of the mechanism of tunicamycin in hibition of IgA and IgE secretion by plasma cells.
    J Biol Chem. 1977 Jun 25;252(12):4402-8 PMID: 325006
  3. Analysis of precursors to the envelope glycoproteins of avian RNA tumor viruses in chicken and quail cells.
    J Virol. 1977 Jun;22(3):598-607 PMID: 195080
  4. Impaired intracellular migration and altered solubility of nonglycosylated glycoproteins of vesicular stomatitis virus and Sindbis virus.
    J Biol Chem. 1977 Dec 25;252(24):9018-23 PMID: 200626
  5. Intracellular precursors to the major glycoprotein of avian oncoviruses in chicken embryo fibroblasts.
    J Virol. 1978 Mar;25(3):845-51 PMID: 205673
  6. Synthesis and infectivity of vesicular stomatitis virus containing nonglycosylated G protein.
    Cell. 1978 Apr;13(4):671-9 PMID: 207437
  7. Proteins of Rous-associated virus 61, an avian retrovirus: common precursor for glycoproteins gp85 and gp35 and use of pactamycin to map translational order of proteins in the gag, pol, and env genes.
    J Virol. 1978 May;26(2):380-8 PMID: 77910
  8. Synthesis and processing of avian sarcoma virus glycoproteins.
    Virology. 1978 Apr;85(2):475-86 PMID: 208237
  9. Effect of tunicamycin on the secretion of serum proteins by primary cultures of rat and chick hepatocytes. Studies on transferrin, very low density lipoprotein, and serum albumin.
    J Biol Chem. 1978 Aug 10;253(15):5332-7 PMID: 209036
  10. alpha1-Antitrypsin: the presence of excess mannose in the Z variant isolated from liver.
    Science. 1978 Sep 29;201(4362):1229-32 PMID: 308696
  11. The structure of the Rous sarcoma virus glycoprotein complex.
    Arch Virol. 1978;58(1):61-4 PMID: 211992
  12. Characterization of the env gene in avian oncoviruses by heteroduplex mapping.
    J Virol. 1978 Sep;27(3):667-76 PMID: 212588
  13. The nonglycosylated glycoprotein of vesicular stomatitis virus is temperature-sensitive and undergoes intracellular aggregation at elevated temperatures.
    J Biol Chem. 1979 May 10;254(9):3600-7 PMID: 218976
  14. Oligosaccharide chains of avian RNA tumor virus glycoproteins contain heterogeneous oligomannosyl cores.
    J Virol. 1979 Jan;29(1):336-43 PMID: 219228
  15. Polarity of influenza and vesicular stomatitis virus maturation in MDCK cells: lack of a requirement for glycosylation of viral glycoproteins.
    Proc Natl Acad Sci U S A. 1979 Dec;76(12):6430-4 PMID: 230510
  16. Relation of fatty acid attachment to the translation and maturation of vesicular stomatitis and Sindbis virus membrane glycoproteins.
    J Biol Chem. 1980 Apr 25;255(8):3334-9 PMID: 6245077
  17. Biosynthesis of lysosomal enzymes in fibroblasts. Phosphorylation of mannose residues.
    J Biol Chem. 1980 May 25;255(10):4946-50 PMID: 6989822
  18. Subcellular compartmentalization of saccharide moieties in cultured normal and malignant cells.
    J Cell Biol. 1980 May;85(2):429-34 PMID: 7372714
  19. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  20. Structural mutations in a mouse immunoglobulin light chain resulting in failure to be secreted.
    Cell. 1980 Jun;20(2):283-92 PMID: 6156005
  21. Mutants of vesicular stomatitis virus blocked at different stages in maturation of the viral glycoprotein.
    Cell. 1980 Sep;21(2):417-27 PMID: 6250721
  22. Deletion mutants of SV40 that affect the structure of viral tumor antigens.
    Cold Spring Harb Symp Quant Biol. 1980;44 Pt 1,:285-91 PMID: 6253142
  23. Structure and processing of the mouse mammary tumor virus glycoprotein precursor pr73env.
    J Virol. 1980 Aug;35(2):349-61 PMID: 6255176
  24. Intracellular transport of secretory and membrane proteins in hepatoma cells infected by vesicular stomatitis virus.
    Cell. 1980 Dec;22(3):709-17 PMID: 6257395
  25. Reversible defect in the glycosylation of the membrane proteins of Semliki Forest virus ts-1 mutant.
    Virology. 1981 Feb;109(1):165-73 PMID: 7467130
  26. Passage of an integral membrane protein, the vesicular stomatitis virus glycoprotein, through the Golgi apparatus en route to the plasma membrane.
    Proc Natl Acad Sci U S A. 1981 Mar;78(3):1746-50 PMID: 6262824
  27. Glycosylation does not determine segregation of viral envelope proteins in the plasma membrane of epithelial cells.
    J Cell Biol. 1981 May;89(2):230-9 PMID: 6265461
  28. Synthesis and processing of asparagine-linked oligosaccharides.
    Annu Rev Biochem. 1981;50:555-83 PMID: 7023366
  29. Rous sarcoma virus glycoproteins contain hybrid-type oligosaccharides.
    J Virol. 1981 Aug;39(2):646-50 PMID: 6268850
  30. Cell-surface expression of influenza haemagglutinin from a cloned DNA copy of the RNA gene.
    Nature. 1981 Oct 22;293(5834):620-5 PMID: 6270568
  31. Mechanisms for the incorporation of proteins in membranes and organelles.
    J Cell Biol. 1982 Jan;92(1):1-22 PMID: 7035466
  32. Two distinct intracellular pathways transport secretory and membrane glycoproteins to the surface of pituitary tumor cells.
    Cell. 1982 Jan;28(1):51-9 PMID: 6279313
  33. Oligosaccharide modifications and the site of processing of gPr92env, the precursor for the viral glycoproteins of Rous sarcoma virus.
    Virology. 1982 May;119(1):122-32 PMID: 6280380
  34. Restriction endonuclease and nucleotide sequence analyses of molecularly cloned unintegrated avian tumor virus DNA: structure of large terminal repeats in circle junctions.
    J Virol. 1982 Apr;42(1):346-51 PMID: 6283156
  35. Sequence of the long terminal repeat and adjacent segments of the endogenous avian virus Rous-associated virus 0.
    J Virol. 1982 Jul;43(1):191-200 PMID: 6286997
  36. Cell surface expression of the influenza virus hemagglutinin requires the hydrophobic carboxy-terminal sequences.
    Cell. 1982 Sep;30(2):649-56 PMID: 6814764
  37. Expression from cloned cDNA of cell-surface secreted forms of the glycoprotein of vesicular stomatitis virus in eucaryotic cells.
    Cell. 1982 Oct;30(3):753-62 PMID: 6291783
  38. Evidence for a glycoprotein "signal" involved in transport between subcellular organelles. Two membrane glycoproteins encoded by murine leukemia virus reach the cell surface at different rates.
    J Biol Chem. 1982 Dec 10;257(23):14011-7 PMID: 7142193
  39. Construction of influenza haemagglutinin genes that code for intracellular and secreted forms of the protein.
    Nature. 1982 Dec 16;300(5893):598-603 PMID: 7144911
  40. Site-specific base substitution and deletion mutations that enhance or suppress transcription of the SV40 major late RNA.
    Cell. 1982 Dec;31(3 Pt 2):625-33 PMID: 7159930
  41. Kinetics of serum protein secretion by cultured hepatoma cells. Evidence for multiple secretory pathways.
    J Biol Chem. 1983 Mar 10;258(5):3304-8 PMID: 6186665
  42. High efficiency polyoma DNA transfection of chloroquine treated cells.
    Nucleic Acids Res. 1983 Mar 11;11(5):1295-308 PMID: 6298741
  43. Nucleotide sequence of Rous sarcoma virus.
    Cell. 1983 Mar;32(3):853-69 PMID: 6299578
  44. Mutational analysis of simian virus 40 T antigen: isolation and characterization of mutants with deletions in the T-antigen gene.
    Mol Cell Biol. 1983 Feb;3(2):203-13 PMID: 6300656
  45. Reversible block in intracellular transport and budding of mutant vesicular stomatitis virus glycoproteins.
    Virology. 1983 Mar;125(2):335-48 PMID: 6301145
  46. Complete sequence of the Rous sarcoma virus env gene: identification of structural and functional regions of its product.
    J Virol. 1983 Jun;46(3):920-36 PMID: 6304351
  47. Analysis of the hemagglutinin glycoprotein from mutants of vaccinia virus that accumulates on the nuclear envelope.
    Cell. 1983 Jun;33(2):423-34 PMID: 6861204
  48. Swainsonine causes the production of hybrid glycoproteins by human skin fibroblasts and rat liver Golgi preparations.
    J Biol Chem. 1983 Jun 25;258(12):7578-85 PMID: 6408079
  49. Hepatoma secretory proteins migrate from rough endoplasmic reticulum to Golgi at characteristic rates.
    Nature. 1983 Jul 7-13;304(5921):80-3 PMID: 6866094
  50. Expression of Semliki Forest virus proteins from cloned complementary DNA. II. The membrane-spanning glycoprotein E2 is transported to the cell surface without its normal cytoplasmic domain.
    J Cell Biol. 1983 Sep;97(3):652-8 PMID: 6688424
  51. Altered cytoplasmic domains affect intracellular transport of the vesicular stomatitis virus glycoprotein.
    Cell. 1983 Sep;34(2):513-24 PMID: 6352053
  52. Expression and function of transplantation antigens with altered or deleted cytoplasmic domains.
    Cell. 1983 Sep;34(2):535-44 PMID: 6604582
  53. Isolation of stable mouse cell lines that express cell surface and secreted forms of the vesicular stomatitis virus glycoprotein.
    J Cell Biol. 1983 Nov;97(5 Pt 1):1381-8 PMID: 6415065
  54. Mechanisms of protein localization.
    Microbiol Rev. 1983 Sep;47(3):313-44 PMID: 6355805
  55. Immunoelectron microscopic studies of the intracellular transport of the membrane glycoprotein (G) of vesicular stomatitis virus in infected Chinese hamster ovary cells.
    J Cell Biol. 1983 Dec;97(6):1777-87 PMID: 6315743
  56. Vesicular stomatitis virus glycoprotein, albumin, and transferrin are transported to the cell surface via the same Golgi vesicles.
    J Cell Biol. 1983 Dec;97(6):1815-22 PMID: 6315744
  57. Construction, expression and recognition of an H-2 molecule lacking its carboxyl terminus.
    Nature. 1984 Feb 2-8;307(5950):432-6 PMID: 6363940
  58. Processing of gPr92env, the precursor to the glycoproteins of Rous sarcoma virus: use of inhibitors of oligosaccharide trimming and glycoprotein transport.
    Virology. 1984 Jan 15;132(1):95-109 PMID: 6320537
  59. Alterations in the transport and processing of Rous sarcoma virus envelope glycoproteins mutated in the signal and anchor regions.
    J Cell Biochem. 1983;23(1-4):81-94 PMID: 6327741
  60. Secretion of a lambda 2 immunoglobulin chain is prevented by a single amino acid substitution in its variable region.
    Cell. 1983 May;33(1):77-83 PMID: 6432336
  61. Enchancement of the infectivity of simian virus 40 deoxyribonucleic acid with diethylaminoethyl-dextran.
    J Natl Cancer Inst. 1968 Aug;41(2):351-7 PMID: 4299537
  62. Subunit structure and number of combining sites of the immunoglobulin A myeloma protein produced by mouse plasmacytoma MOPC-315.
    Biochemistry. 1971 Nov 23;10(24):4359-68 PMID: 5142616
  63. Purification and properties of an endo-beta-N-acetylglucosaminidase from Streptomyces griseus.
    J Biol Chem. 1974 Feb 10;249(3):811-7 PMID: 4204552
  64. Mapping of biological functions on RNA of avian tumor viruses: location of regions required for transformation and determination of host range.
    Proc Natl Acad Sci U S A. 1975 Dec;72(12):4772-6 PMID: 174075
  65. Molecular abnormality of human alpha1-antitrypsin variant (Pi-ZZ) associated with plasma activity deficiency.
    Proc Natl Acad Sci U S A. 1976 Apr;73(4):1324-8 PMID: 1083527
  66. Conversion of beta-galactosidase to a membrane-bound state by gene fusion.
    Proc Natl Acad Sci U S A. 1976 Oct;73(10):3423-7 PMID: 790385
  67. Evidence that a precursor glycoprotein is cleaved to yield the major glycoprotein of avian tumor virus.
    J Virol. 1977 Feb;21(2):810-4 PMID: 189094
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.99.6.2011
Published
1984-12-00
Pages
2011-23
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2113559
Subset
IM
Grants
NCI NIH HHS · CA-00685 · United States
NCI NIH HHS · CA-07396 · United States
NCI NIH HHS · CA-29884 · United States
External Links
PubMed source DOI: 10.1083/jcb.99.6.2011 PMC Full Text View journal details
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