Colocalization of adeno-associated virus Rep and capsid proteins in the nuclei of infected cells
Open accessAbstract
The mechanism of adeno-associated virus (AAV) DNA replication was characterized both genetically and biochemically. In this study, we used monoclonal and polyclonal antibodies to examine the AAV p5 (Rep78 and Rep68) and p19 (Rep52 and Rep40) proteins in infected cells. By overexpressing a truncated Rep78 protein in Escherichia coli, we obtained monoclonal antibody anti-78/68, which is specific for the p5 Rep proteins, and monoclonal antibody anti-52/40, which recognized both the p5 and p19 Rep proteins. In single-fluorochrome indirect immunofluorescence labeling experiments, the viral Rep proteins were localized in distinct intranuclear foci. Analysis of AAV proteins by double-fluorochrome indirect immunofluorescence experiments demonstrated that (i) all four AAV Rep proteins occupied the same intranuclear compartments and (ii) the Rep and capsid proteins colocalized in the nuclei of infected cells. These results suggest that replication centers similar to those established by other viruses exist for AAV. These reagents should provide a useful tool for further delineation of the mechanism of AAV replication in vitro.
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Capsids colocalize with both Rep proteins and the ssDNA.
“This is followed by the observation of capsids colocalizing with both Rep proteins and the ssDNA [26, 41–42].”
From Understanding Capsid Assembly and Genome Packaging for Adeno-Associated Viruses (2017) · cites this paper as [41]
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AAV Capsid Structure and Cell Interactions
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Understanding Capsid Assembly and Genome Packaging for Adeno-Associated Viruses
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Development of AAV serotype-specific ELISAs using novel monoclonal antibodies
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- Authors
- Hunter, L. A., & Samulski, R. J.
- Affiliations
- University of Pittsburgh (US)
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 1992-01
- Volume
- 66
- Issue
- 1
- Pages
- 317-324
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- PMC
- PMC238290
- OpenAlex
- W1568306217
- Cited by
- 68
- References
- 1
- Citations by year
- 2023: 3 · 2022: 1 · 2021: 3 · 2018: 1 · 2017: 1 · 2016: 1 · 2014: 1 · 2013: 2
- FWCI
- 1.31
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Virology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r29ai025530-02)
- National Institute of Allergy and Infectious Diseases (AI 25530-03)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Health Sciences › Medicine › Infectious Diseases › Parvovirus B19 Infection Studies
- Physical Sciences › Environmental Science › Ecology › Bacteriophages and microbial interactions
- Keywords
- adeno-associated virus, Rep protein, Rep78, Rep68, capsid protein, protein colocalization, viral replication centers, monoclonal antibodies, indirect immunofluorescence
- MeSH terms
- Dependovirus, Antibodies, Monoclonal, Epitopes, Capsid, Cell Nucleus, Cells, Cultured, Cloning, Molecular, DNA-Binding Proteins, Escherichia coli, Fluorescent Antibody Technique, Humans, Viral Proteins, Immunoblotting
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