Properties of the Adeno-Associated Virus Assembly-Activating Protein
Open accessAbstract
Adeno-associated virus (AAV) capsid assembly requires expression of the assembly-activating protein (AAP) together with capsid proteins VP1, VP2, and VP3. AAP is encoded by an alternative open reading frame of the cap gene. Sequence analysis and site-directed mutagenesis revealed that AAP contains two hydrophobic domains in the N-terminal part of the molecule that are essential for its assembly-promoting activity. Mutation of these sequences reduced the interaction of AAP with the capsid proteins. Deletions and a point mutation in the capsid protein C terminus also abolished capsid assembly and strongly reduced the interaction with AAP. Interpretation of these observations on a structural basis suggests an interaction of AAP with the VP C terminus, which forms the capsid protein interface at the 2-fold symmetry axis. This interpretation is supported by a decrease in the interaction of monoclonal antibody B1 with VP3 under nondenaturing conditions in the presence of AAP, indicative of steric hindrance of B1 binding to its C-terminal epitope by AAP. In addition, AAP forms high-molecular-weight oligomers and changes the conformation of nonassembled VP molecules as detected by conformation-sensitive monoclonal antibodies A20 and C37. Combined, these observations suggest a possible scaffolding activity of AAP in the AAV capsid assembly reaction.
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All serotypes tested, with the exception of AAV4, AAV5 and AAV11, require AAP in order to assemble their capsid.
“In the most recent studies, it has been shown that all serotypes tested, with the exception of AAV4, AAV5 and AAV11, require AAP in order to assemble their capsid [35,39, 45–47].”
From Understanding Capsid Assembly and Genome Packaging for Adeno-Associated Viruses (2017) · cites this paper as [39]
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“The application of MAbs A20 and C37 showed similar reactions in the intermediate-size fraction and the 60S fractions but revealed, in addition, a significant reaction with VP3 in the monomer/oligomer fractions at around 6.5S when AAP was coexpressed.”
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- Authors
- Naumer, M., Sonntag, F., Schmidt, K., Nieto, K., Panke, C., Davey, N. E., Popa-Wagner, R., & Kleinschmidt, J. A.
- Affiliations
- German Cancer Research Center (DE)
- European Molecular Biology Laboratory (DE)
- ORCID
- Davey, Norman E.
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 2012-12
- Volume
- 86
- Issue
- 23
- Pages
- 13038-13048
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- PMC
- PMC3497687
- OpenAlex
- W2160630690
- Cited by
- 97
- References
- 8
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 6 · 2023: 7 · 2022: 2 · 2021: 16 · 2020: 5 · 2019: 5 · 2018: 10 · 2017: 11 · 2016: 5 · 2015: 8 · 2014: 6 · 2013: 9
- FWCI
- 4.43
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Journal of Virology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Deutsche Forschungsgemeinschaft
- Deutsche Krebshilfe
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Physical Sciences › Environmental Science › Ecology › Bacteriophages and microbial interactions
- Health Sciences › Medicine › Epidemiology › Virology and Viral Diseases
- Keywords
- adeno-associated virus, capsid assembly, VP1, VP2, VP3, hydrophobic domains, protein-protein interaction, protein oligomerization, monoclonal antibodies, alternative reading frame
- MeSH terms
- Dependovirus, Antibodies, Monoclonal, Capsid, Microscopy, Electron, Immunoblotting, Mutagenesis, Site-Directed, Sequence Analysis, DNA, Protein Structure, Tertiary, Virus Assembly, Capsid Proteins, Immunoprecipitation
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