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PMID: 2109101 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Assembly of gag-beta-galactosidase proteins into retrovirus particles.

Journal of virology ·Vol. 64 ·No. 5 ·1990-05-00 ·Pages 2265-79

Jones TA, Blaug G, Hansen M, Barklis E

Abstract

We studied the expression of beta-galactosidase (beta-gal) and 15 gag-beta-gal fusion proteins in the presence of Moloney murine leukemia virus wild-type core (gag) proteins. Analysis indicated that proteins retaining the amino-terminal portion of gag through the capsid protein-coding region were incorporated into retrovirus particles. Proteins which deleted portions of the capsid protein were assembled into virions at low efficiency, indicating the importance of capsid protein interactions in retrovirus assembly. Fusion proteins which retained the amino-terminal matrix protein of the gag polyprotein but which lacked the capsid protein were released efficiently from cells in a nonviral form. The nonviral form was characterized by a high sedimentation coefficient and a low density, suggestive of membrane vesicles. While beta-gal was present in the cytoplasm of expressing cells, all fusion constructs were associated with cellular membranes. gag-beta-gal proteins which were capable of release from cells demonstrated a two-component immunofluorescence staining pattern consisting of a circle of fluorescence around the nucleus and a punctate pattern of staining throughout the remainder of the cell. Interestingly, fusions within the matrix protein were trapped intracellularly and yielded distinct perinuclear staining patterns, possibly localizing to the rough endoplasmic reticulum and/or Golgi. This observation suggests that Moloney murine leukemia virus gag proteins travel to the plasma membrane by vesicular transport associated with the cytoplasmic face of intracellular vesicles.

MeSH Terms
Animals Base Sequence Capsid/genetics Cell Line Cell Membrane/metabolism Cells, Cultured Chromosome Deletion Codon/genetics Fluorescent Antibody Technique Galactosidases/genetics Genes, gag Models, Biological Molecular Sequence Data Moloney murine leukemia virus/genetics Recombinant Fusion Proteins/biosynthesis,isolation & purification Retroviridae/genetics Viral Envelope Proteins/biosynthesis,genetics,isolation & purification beta-Galactosidase/biosynthesis,genetics,isolation & purification
Chemicals
Codon Recombinant Fusion Proteins Viral Envelope Proteins Galactosidases beta-Galactosidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Jones T A
Vollum Institute for Advanced Biomedical Research, Portland, Oregon.
Blaug G
Hansen M
Barklis E
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.64.5.2265-2279.1990
Published
1990-05-00
Pages
2265-79
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC249388
Subset
IM
Grants
NCI NIH HHS · 5R01 CA47088-02 · United States
External Links
PubMed source DOI: 10.1128/JVI.64.5.2265-2279.1990 PMC Full Text View journal details
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