Assembly of viruslike particles by recombinant structural proteins of adeno-associated virus type 2 in insect cells
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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.
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Mutational studies on the entire cap gene showed that the VP3 common sequence is enough to assemble AAV capsids by knocking out the start codons of VP1 and VP2.
“Mutational studies on the entire cap gene showed that the VP3 common sequence is enough to assemble AAV capsids by knocking out the start codons of VP1 and VP2 [31–34].”
From Understanding Capsid Assembly and Genome Packaging for Adeno-Associated Viruses (2017) · cites this paper as [31]
Cites
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- Authors
- Ruffing, M., Zentgraf, H., & Kleinschmidt, J. A.
- Affiliations
- German Cancer Research Center (DE)
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 1992-12
- Volume
- 66
- Issue
- 12
- Pages
- 6922-6930
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- PMC
- PMC240317
- OpenAlex
- W1500537600
- Cited by
- 109
- References
- 1
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 3 · 2023: 1 · 2021: 2 · 2020: 2 · 2019: 3 · 2018: 3 · 2017: 3 · 2016: 3 · 2015: 2 · 2014: 2 · 2012: 2
- FWCI
- 1.59
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Virology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Health Sciences › Medicine › Epidemiology › Virology and Viral Diseases
- Keywords
- adeno-associated virus type 2, virus-like particles, capsid protein, VP1, VP2, VP3, recombinant baculovirus expression, insect cells, Sf9 cells, HeLa cells, empty capsids, capsid assembly, nuclear localization, overlapping open reading frames
- MeSH terms
- Dependovirus, Animals, Capsid, Cell Line, Cell Nucleus, HeLa Cells, Humans, Insecta, Microscopy, Electron, Plasmids, Restriction Mapping, Viral Structural Proteins, Gene Expression, Mutagenesis, Site-Directed, Open Reading Frames, Genome, Viral
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