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PMID: 3142146 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Intracellular accumulation of Punta Toro virus glycoproteins expressed from cloned cDNA.

Virology ·Vol. 167 ·No. 1 ·1988-11-00 ·Pages 251-60

Matsuoka Y, Ihara T, Bishop DH, Compans RW

Abstract

The Punta Toro virus (PTV) middle size (M) RNA encodes two glycoproteins, G1 and G2, and possibly a nonstructural protein, NSM. A partial cDNA clone of the M segment which contains G1 and G2 glycoprotein coding sequences but lacks most of the NSM sequences was inserted into the genome of vaccinia virus under the control of an early vaccinia promoter. Cells infected with the recombinant virus were found to synthesize two polypeptides with molecular weights of 65,000 (G1) and 55,000 (G2) that reacted specifically with antibody against PTV. Studies using indirect immunofluorescence microscopy revealed that these proteins accumulated intracellularly in the perinuclear region. The results of endoglycosidase H digestion of these glycoproteins suggested that both G1 and G2 glycoproteins were transported from the RER to the Golgi complex. These proteins were not chased out from the Golgi region during a 6-hr incubation in the presence of cycloheximide. Surface immune precipitation and 125I-protein A binding assays also demonstrated that the majority of the G1 and G2 glycoproteins are retained intracellularly. These results indicate that the PTV glycoproteins contain the necessary information for retention in the Golgi apparatus.

MeSH Terms
Acetylglucosaminidase/metabolism Animals Bunyaviridae/genetics Cloning, Molecular DNA, Viral/genetics Electrophoresis, Polyacrylamide Gel Fluorescent Antibody Technique Gene Expression Regulation Glycoproteins/analysis,biosynthesis,genetics HeLa Cells Hemagglutinins, Viral/biosynthesis,genetics Humans Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase Plasmids Precipitin Tests Tunicamycin/pharmacology Vaccinia virus/genetics Vero Cells Viral Proteins/analysis,biosynthesis,genetics
Chemicals
DNA, Viral Glycoproteins Hemagglutinins, Viral Viral Proteins Tunicamycin Acetylglucosaminidase Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Matsuoka Y
Department of Microbiology, University of Alabama, Birmingham 35294.
Ihara T
Bishop D H
Compans R W
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Article Info
Journal
Virology
Abbr.
Virology
ISSN
0042-6822
DOI
10.1016/0042-6822(88)90075-x
Published
1988-11-00
Pages
251-60
Language
English
Region
United States
NLM ID
0110674
PMCID
PMC7130723
Subset
IM
Grants
NIAID NIH HHS · AI 12680 · United States
External Links
PubMed source DOI: 10.1016/0042-6822(88)90075-x PMC Full Text View journal details
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