The Assembly-Activating Protein Promotes Capsid Assembly of Different Adeno-Associated Virus Serotypes
Open accessAbstract
Adeno-associated virus type 2 (AAV2) capsid assembly requires the expression of a virally encoded assembly-activating protein (AAP). By providing AAP together with the capsid protein VP3, capsids are formed that are composed of VP3 only. Electron cryomicroscopy analysis of assembled VP3-only capsids revealed all characteristics of the wild-type AAV2 capsids. However, in contrast to capsids assembled from VP1, VP2, and VP3, the pores of VP3-only capsids were more restricted at the inside of the 5-fold symmetry axes, and globules could not be detected below the 2-fold symmetry axes. By comparing the capsid assembly of several AAV serotypes with AAP protein from AAV2 (AAP-2), we show that AAP-2 is able to efficiently stimulate capsid formation of VP3 derived from several serotypes, as demonstrated for AAV1, AAV2, AAV8, and AAV9. Capsid formation, by coexpressing AAV1-, AAV2-, or AAV5-VP3 with AAP-1, AAP-2, or AAP-5 revealed the ability of AAP-1 and AAP-2 to complement each other in AAV1 and AAV2 assembly, whereas for AAV5 assembly more specific conditions are required. Sequence alignment of predicted AAP proteins from the known AAV serotypes indicates a high degree of homology of all serotypes to AAP-2 with some divergence for AAP-4, AAP-5, AAP-11, and AAP-12. Immunolocalization of assembled capsids from different serotypes confirmed the preferred nucleolar localization of capsids, as observed for AAV2; however, AAV8 and AAV9 capsids could also be detected throughout the nucleus. Taken together, the data show that AAV capsid assembly of different AAV serotypes also requires the assistance of AAP proteins.
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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 [46]
The paper’s Results report that AAV5-VP3 capsid formation was restored by AAP-5, and its summary explicitly says AAV5 assembly required AAP-5. Thus AAV5 is not an exception to AAP dependence in this source. The paper discusses AAV4 and AAV11 sequence differences but does not test their assembly requirement. (LLM, conf=0.92)
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Cites
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Self-citation
- Authors
- Sonntag, F., Köther, K., Schmidt, K., Weghofer, M., Raupp, C., Nieto, K., Kuck, A., Gerlach, B., Böttcher, B., Müller, O. J., Lux, K., Hörer, M., & Kleinschmidt, J. A.
- Affiliations
- German Cancer Research Center (DE)
- Medigene (Germany) (DE)
- University of Edinburgh (GB)
- Heidelberg University (DE)
- Rentschler Biopharma (Germany) (DE)
- Corresponding author
- Kleinschmidt, J. A.
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 2011-12
- Volume
- 85
- Issue
- 23
- Pages
- 12686-12697
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- PMC
- PMC3209379
- OpenAlex
- W2098880579
- Cited by
- 184
- References
- 9
- Citations by year
- 2026: 8 · 2025: 10 · 2024: 20 · 2023: 12 · 2022: 12 · 2021: 20 · 2020: 14 · 2019: 13 · 2018: 20 · 2017: 11 · 2016: 7 · 2015: 7 · 2014: 8 · 2013: 8 · 2012: 11
- FWCI
- 5.59
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Virology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Edinburgh Research Explorer (University of Edinburgh) (submittedVersion, other-oa)
- Funders
- Deutsche Forschungsgemeinschaft
- Universität Heidelberg
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Environmental Science › Ecology › Bacteriophages and microbial interactions
- Keywords
- adeno-associated virus, capsid assembly, AAV serotypes, VP3 capsid protein, AAV1, AAV2, AAV8, AAV9, AAV5, capsid structure, nucleolar localization, protein complementation
- MeSH terms
- Dependovirus, Animals, Antibodies, Monoclonal, Capsid, Cells, Cultured, Female, Fluorescent Antibody Technique, HeLa Cells, Humans, Kidney, Mice, Inbred C57BL, RNA, Messenger, Serotyping, Virion, Blotting, Western, Virus Assembly, Capsid Proteins, Mice, Real-Time Polymerase Chain Reaction
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