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PMID: 1985194 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Internally located cleavable signal sequences direct the formation of Semliki Forest virus membrane proteins from a polyprotein precursor.

Journal of virology ·Vol. 65 ·No. 1 ·1991-01-00 ·Pages 147-54

Liljeström P, Garoff H

Abstract

The proteolytic processes involved in the cotranslational production of the Semliki Forest virus proteins p62, 6K, and E1 from a common precursor polypeptide were analyzed by an in vitro translation-translocation assay. By studying the behavior of wild-type and mutant variants of the polyprotein, we show that the signal sequences responsible for membrane translocation of the 6K and E1 proteins reside in the C-terminal regions of p62 and 6K, respectively. We present evidence suggesting that the polyprotein is processed on the luminal side by signal peptidase at consensus cleavage sites immediately following the signal sequences. Our results also lead us to conclude that the 6K protein is a transmembrane polypeptide with its N terminus on the luminal side of the membrane (type I). Thus, the production of all three membrane proteins is directed by alternating signal and stop-transfer (anchor) sequences that function in translocation and cleavage of the virus precursor polyprotein. This also shows conclusively that internally located signal sequences can be cleaved by signal peptidase.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Chromosome Deletion DNA, Viral/genetics Escherichia coli/genetics Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Oligonucleotide Probes Protein Biosynthesis Protein Conformation Protein Precursors/genetics Protein Processing, Post-Translational Protein Sorting Signals/genetics,metabolism Rabbits Restriction Mapping Reticulocytes/metabolism Semliki forest virus/genetics Viral Matrix Proteins/genetics Viral Structural Proteins/genetics
Chemicals
DNA, Viral Oligonucleotide Probes Protein Precursors Protein Sorting Signals Viral Matrix Proteins Viral Structural Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Liljeström P
Department of Molecular Biology, Karolinska Institute, Huddinge, Sweden.
Garoff H
References (40)
40 references, click to expand
  1. Protein oligomerization in the endoplasmic reticulum.
    Annu Rev Cell Biol. 1989;5:277-307 PMID: 2688707
  2. Reinitiation of translocation in the Semliki Forest virus structural polyprotein: identification of the signal for the E1 glycoprotein.
    EMBO J. 1986 Jul;5(7):1551-60 PMID: 3017702
  3. Truncations of a secretory protein define minimum lengths required for binding to signal recognition particle and translocation across the endoplasmic reticulum membrane.
    J Biol Chem. 1990 May 5;265(13):7478-84 PMID: 2185250
  4. Site-directed mutagenesis of the proposed catalytic amino acids of the Sindbis virus capsid protein autoprotease.
    J Virol. 1990 Jun;64(6):3069-73 PMID: 2335827
  5. Assembly of the Semliki Forest virus membrane glycoproteins in the membrane of the endoplasmic reticulum in vitro.
    J Mol Biol. 1978 Oct 5;124(4):587-600 PMID: 712848
  6. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
    Gene. 1977;2(2):95-113 PMID: 344137
  7. The signal sequence of the p62 protein of Semliki Forest virus is involved in initiation but not in completing chain translocation.
    J Cell Biol. 1990 Sep;111(3):867-76 PMID: 2391367
  8. Evidence for an autoprotease activity of sindbis virus capsid protein.
    Virology. 1978 Oct 15;90(2):366-9 PMID: 726255
  9. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
    Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
  10. Complete structure of the hemagglutinin gene from the human influenza A/Victoria/3/75 (H3N2) strain as determined from cloned DNA.
    Cell. 1980 Mar;19(3):683-96 PMID: 6153930
  11. Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate.
    Anal Biochem. 1979 Sep 15;98(1):132-5 PMID: 543547
  12. Carboxyl-terminal sequence analysis of the four structural proteins of Semliki Forest virus.
    FEBS Lett. 1980 Jun 30;115(2):163-6 PMID: 7398872
  13. Nucleotide sequence of cdna coding for Semliki Forest virus membrane glycoproteins.
    Nature. 1980 Nov 20;288(5788):236-41 PMID: 6985476
  14. Vesicular stomatitis virus glycoprotein is anchored in the viral membrane by a hydrophobic domain near the COOH terminus.
    Proc Natl Acad Sci U S A. 1980 Jul;77(7):3884-8 PMID: 6253998
  15. Structure and assembly of alphaviruses.
    Curr Top Microbiol Immunol. 1982;99:1-50 PMID: 6284443
  16. 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
  17. Ross River virus 26 s RNA: complete nucleotide sequence and deduced sequence of the encoded structural proteins.
    Virology. 1983 Aug;129(1):170-87 PMID: 6310876
  18. Preparation and use of nuclease-treated rabbit reticulocyte lysates for the translation of eukaryotic messenger RNA.
    Methods Enzymol. 1983;96:50-74 PMID: 6656641
  19. Dog pancreatic microsomal-membrane polypeptides analysed by two-dimensional gel electrophoresis.
    Biochem J. 1984 Jan 1;217(1):145-57 PMID: 6696719
  20. Glycosylation and surface expression of the influenza virus neuraminidase requires the N-terminal hydrophobic region.
    Mol Cell Biol. 1984 Jan;4(1):8-16 PMID: 6700587
  21. Complete nucleotide sequence of the genomic RNA of Sindbis virus.
    Virology. 1984 Feb;133(1):92-110 PMID: 6322438
  22. Processing of the Semliki Forest virus structural polyprotein: role of the capsid protease.
    J Virol. 1987 May;61(5):1301-9 PMID: 3553612
  23. Nucleotide sequence of the genome region encoding the 26S mRNA of eastern equine encephalomyelitis virus and the deduced amino acid sequence of the viral structural proteins.
    J Gen Virol. 1987 Aug;68 ( Pt 8):2129-42 PMID: 2886548
  24. The structure and function of the hemagglutinin membrane glycoprotein of influenza virus.
    Annu Rev Biochem. 1987;56:365-94 PMID: 3304138
  25. A specific transmembrane domain of a coronavirus E1 glycoprotein is required for its retention in the Golgi region.
    J Cell Biol. 1987 Sep;105(3):1205-14 PMID: 2821010
  26. In vitro RNA synthesis with SP6 RNA polymerase.
    Methods Enzymol. 1987;155:397-415 PMID: 2828872
  27. Transcending the impenetrable: how proteins come to terms with membranes.
    Biochim Biophys Acta. 1988 Jun 9;947(2):307-33 PMID: 3285892
  28. A rapid and convenient method for the preparation and storage of competent bacterial cells.
    Nucleic Acids Res. 1988 Apr 25;16(8):3580 PMID: 3287331
  29. Western equine encephalitis virus is a recombinant virus.
    Proc Natl Acad Sci U S A. 1988 Aug;85(16):5997-6001 PMID: 3413072
  30. A multisite-directed mutagenesis using T7 DNA polymerase: application for reconstructing a mammalian gene.
    Gene. 1988 Sep 15;69(1):81-9 PMID: 3224824
  31. Signal sequences.
    Biochemistry. 1989 Feb 7;28(3):923-30 PMID: 2653440
  32. The full-length nucleotide sequences of the virulent Trinidad donkey strain of Venezuelan equine encephalitis virus and its attenuated vaccine derivative, strain TC-83.
    Virology. 1989 May;170(1):19-30 PMID: 2524126
  33. Sequence and topology of a model intracellular membrane protein, E1 glycoprotein, from a coronavirus.
    Nature. 1984 Apr 19-25;308(5961):751-2 PMID: 6325918
  34. NH2-terminal hydrophobic region of influenza virus neuraminidase provides the signal function in translocation.
    Proc Natl Acad Sci U S A. 1984 Apr;81(8):2327-31 PMID: 6326121
  35. Role of signal recognition particle in the membrane assembly of Sindbis viral glycoproteins.
    Eur J Biochem. 1984 May 2;140(3):499-502 PMID: 6723645
  36. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  37. Sequence analysis of three Sindbis virus mutants temperature-sensitive in the capsid protein autoprotease.
    Proc Natl Acad Sci U S A. 1985 Jul;82(14):4648-52 PMID: 3895223
  38. Mutants of the membrane-binding region of Semliki Forest virus E2 protein. I. Cell surface transport and fusogenic activity.
    J Cell Biol. 1986 Mar;102(3):889-901 PMID: 3753980
  39. Mutants of the membrane-binding region of Semliki Forest virus E2 protein. II. Topology and membrane binding.
    J Cell Biol. 1986 Mar;102(3):902-10 PMID: 3949882
  40. Complete sequence of the genomic RNA of O'nyong-nyong virus and its use in the construction of alphavirus phylogenetic trees.
    Virology. 1990 Mar;175(1):110-23 PMID: 2155505
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.65.1.147-154.1991
Published
1991-01-00
Pages
147-54
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC240499
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.65.1.147-154.1991 PMC Full Text View journal details
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