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PMID: 1331503 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Assembly of viruslike particles by recombinant structural proteins of adeno-associated virus type 2 in insect cells.

Journal of virology ·Vol. 66 ·No. 12 ·1992-12-00 ·Pages 6922-30

Ruffing M, Zentgraf H, Kleinschmidt JA

Abstract

The three capsid proteins VP1, VP2, and VP3 of the adeno-associated virus type 2 (AAV-2) are encoded by overlapping sequences of the same open reading frame. Separate expression of these proteins by recombinant baculoviruses in insect cells was achieved by mutation of the internal translation initiation codons. Coexpression of VP1 and VP2, VP2 and VP3, and all three capsid proteins and the expression of VP2 alone in Sf9 cells resulted in the production of viruslike particles resembling empty capsids generated during infection of HeLa cells with AAV-2 and adenovirus. These results suggest a requirement for VP2 in the formation of empty capsids. Individual expression of the AAV capsid proteins in HeLa cells showed that VP1 and VP2 accumulate in the cell nucleus and VP3 is distributed between nucleus and cytoplasm. Coexpression of VP3 with the other structural proteins also led to nuclear localization of VP3, indicating that the formation of a complex with VP1 or VP2 is required for accumulation of VP3 in the nucleus.

MeSH Terms
Animals Capsid/biosynthesis,genetics,ultrastructure Cell Line Cell Nucleus/metabolism,ultrastructure Dependovirus/genetics,physiology Gene Expression Genome, Viral HeLa Cells Humans Insecta Microscopy, Electron Mutagenesis, Site-Directed Open Reading Frames Plasmids Restriction Mapping Viral Structural Proteins/biosynthesis,genetics
Chemicals
Viral Structural Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ruffing M
Deutsches Krebsforschungszentrum, Forschungsschwerpunkt Angewandte Tumorvirologie, Heidelberg, Germany.
Zentgraf H
Kleinschmidt J A
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.66.12.6922-6930.1992
Published
1992-12-00
Pages
6922-30
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC240317
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.66.12.6922-6930.1992 PMC Full Text View journal details
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