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PMID: 3753980 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mutants of the membrane-binding region of Semliki Forest virus E2 protein. I. Cell surface transport and fusogenic activity.

The Journal of cell biology ·Vol. 102 ·No. 3 ·1986-03-00 ·Pages 889-901

Cutler DF, Garoff H

Abstract

Three mutations of the membrane-binding region of the Semliki Forest virus (SFV) p62 polypeptide (the precursor for virion E3 and E2) have been made by oligonucleotide-directed mutagenesis of a cDNA clone encoding the SFV structural proteins. One of the mutations (A2) substitutes a Glu for an Ala in the middle of the hydrophobic stretch which spans the bilayer. A1 and A3 alter the two basic charged amino acids in the cytoplasmic domain next to the hydrophobic region. The wild-type charge cluster of Arg-Ser-Lys (+2) has been changed to Gly-Ser-Met (0;A3) or to Gly-Ser-Glu (-1;A1). The mutant p62 proteins have been analyzed both in the presence and the absence of E1, the other half of the heterodimer spike complex of SFV. The mutant proteins expressed in COS-7 cells are glycosylated and are of the expected sizes. When co-expressed with E1, all three mutants are cleaved to yield the E2 protein and transported to the surface of COS-7 cells. When expressed in the absence of E1, the mutant p62 proteins remain uncleaved but still reach the cell surface. Once at the cell surface, all three mutants, when co-expressed with E1, can promote low pH-triggered cell-cell fusion. These results show that the three mutant p62/E2 proteins are still membrane associated in a functionally unaltered way.

MeSH Terms
Amino Acid Sequence Animals Biological Transport, Active Cell Fusion Cell Line Cell Membrane/metabolism Chlorocebus aethiops Cloning, Molecular DNA/genetics DNA, Recombinant Fibroblasts/metabolism Hydrogen-Ion Concentration Kidney Membrane Proteins/metabolism Protein Processing, Post-Translational Semliki forest virus/genetics,physiology Viral Proteins/genetics,metabolism,physiology
Chemicals
DNA, Recombinant Membrane Proteins Viral Proteins DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Cutler D F
Garoff H
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.102.3.889
Published
1986-03-00
Pages
889-901
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2114105
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.102.3.889 PMC Full Text View journal details
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