Subcellular compartmentalization of adeno-associated virus type 2 assembly

Wistuba, A; Kern, Andrea; Weger, Stefan; Grimm, Dirk; Kleinschmidt, Jürgen A.

doi:10.1128/jvi.71.2.1341-1352.1997

Cited by 181 publications

(111 citation statements)

References 67 publications

(84 reference statements)

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“…Adeno-associated viruses (AAVs) need a helper virus to initiate their viral cycle. In cells coinfected with AAV and adenovirus (AdV), nucleoli increase in size and viral capsid (Cap) and replication (Rep) proteins are localised in nucleoli [49][50][51]. The relevance of Cap/Rep nucleolar targeting is developed in a following paragraph.…”

Section: Traffickingmentioning

confidence: 99%

“…A role for B23 in the assembly of AAV Caps has been well documented recently, even if it has been known for many years that AAV Cap assembly takes place in nucleoli of AAV-AdV coinfected cells [49][50][51]. Indeed, Cap proteins first accumulate in the nucleoli, then spread over the whole nucleoplasm.…”

Section: Virus Assembly and Egressmentioning

confidence: 99%

“…Indeed, Cap proteins first accumulate in the nucleoli, then spread over the whole nucleoplasm. Expression of the Cap gene alone leads to the formation of capsids which are restricted to the nucleoli, while upon coexpression of Rep, Cap proteins leave the nucleolus, suggesting a role for Rep in capsid export [49][50][51]. B23 protein, in addition to its ability to interact with AAV Rep proteins, also stimulates the interactions of Rep with the AAV genome [51].…”

Section: Virus Assembly and Egressmentioning

confidence: 99%

“…B23 protein, in addition to its ability to interact with AAV Rep proteins, also stimulates the interactions of Rep with the AAV genome [51]. A model has been proposed, in which Cap proteins accumulate and assemble in the nucleolus where they associate with B23 and perhaps other nucleolar proteins, and AAV Rep proteins favour the exit of complexes containing Rep, B23 and Cap proteins from the nucleolus to the nucleoplasm where viral DNA encapsidation occurs [49][50][51].…”

Section: Virus Assembly and Egressmentioning

confidence: 99%

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Involvement of the nucleolus in replication of human viruses

Greco

2009

Reviews in Medical Virology

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Viruses are intracellular pathogens that have to usurp some of the cellular machineries to provide an optimal environment for their own replication. An increasing number of reports reveal that many viruses induce modifications of nuclear substructures including nucleoli, whether they replicate or not in the nucleus of infected cells. Indeed, during infection of cells with various types of human viruses, nucleoli undergo important morphological modifications. A large number of viral components traffic to and from the nucleolus where they interact with different cellular and/or viral factors, numerous host nucleolar proteins are redistributed in other cell compartments or are modified and some cellular proteins are delocalized in the nucleolus of infected cells. Well-documented studies have established that several of these nucleolar modifications play a role in some steps of the viral cycle, and also in fundamental cellular pathways. The nucleolus itself is the place where several essential steps of the viral cycle take place. In other cases, viruses divert host nucleolar proteins from their known functions in order to exert new unexpected role(s).

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Section: Traffickingmentioning

confidence: 99%

Section: Virus Assembly and Egressmentioning

confidence: 99%

Section: Virus Assembly and Egressmentioning

confidence: 99%

Section: Virus Assembly and Egressmentioning

confidence: 99%

See 2 more Smart Citations

Involvement of the nucleolus in replication of human viruses

Greco

2009

Reviews in Medical Virology

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“…Assays used for product characterization and safety testing are included in Table 2.3. [192,193] Determine titer of infectious particles produced Serial Dilution Replication Assay (dRA) [194] Vector genome Dot-blot hybridization Determine genome-containing vector concentration PCR [195] Capsid ELISA [24] Determine total capsid protein concentration -enables a determination of empty particles in the prep Bradford Western Electron microscopy [196] Optical density (OD) [197] Purity…”

Section: Vector Characterizationmentioning

confidence: 99%

The Role of the Adeno-Associated Virus Capsid in Gene Transfer

Vliet

Blouin

Brument

et al. 2008

Methods in Molecular Biology™

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Adeno-associated virus (AAV) is one of the most promising viral gene transfer vectors that has been shown to effect long-term gene expression and disease correction with low toxicity in animal models, and is well tolerated in human clinical trials. The surface of the AAV capsid is an essential component that is involved in cell binding, internalization, and trafficking within the targeted cell. Prior to developing a gene therapy strategy that utilizes AAV, the serotype should be carefully considered since each capsid exhibits a unique tissue tropism and transduction efficiency. Several approaches have been undertaken in an effort to target AAV vectors to specific cell types, including utilizing natural serotypes that target a desired cellular receptor, producing pseudotyped vectors, and engineering chimeric and mosaic AAV capsids. These capsid modifications are being incorporated into vector production and purification methods that provide for the ability to scale-up the manufacturing process to support human clinical trials. Protocols for small-scale and large-scale production of AAV, as well as assays to characterize the final vector product, are presented here. The structures of AAV2, AAV4, and AAV5 have been solved by X-ray crystallography or cryo-electron microscopy (cryo-EM), and provide a basis for rational vector design in developing customized capsids for specific targeting of AAV vectors. The capsid of AAV has been shown to be remarkably stable, which is a desirable characteristic for a gene therapy vector; however, recently it has been shown that the AAV serotypes exhibit differential susceptibility to proteases. The capsid fragmentation pattern when exposed to various proteases, as well as the susceptibility of the serotypes to a series of proteases, provides a unique fingerprint for each serotype that can be used for capsid identity validation. In addition to serotype identification, protease susceptibility can also be utilized to study dynamic structural changes that must occur for the AAV capsid to perform its various functions during the virus life cycle. The use of proteases for structural studies in solution complements the crystal structural studies of the virus. A generic protocol based on proteolysis for AAV serotype identification is provided here.

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Update on the prevalence of serum antibodies (IgG and IgM) to adeno-associated virus (AAV)

Erles¹,

Sebökovà²,

Schlehofer³

1999

J. Med. Virol.

312

214

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In view of presumed non-pathogenicity, tumor suppressive properties, and site-specific integration of the viral genome the human parvovirus, adeno-associated virus (AAV) has gained great interest as a gene transduction vector. Data on the seroprevalence of antibodies to AAV vary between reports, probably due to the different serological methods used. In order to understand better the immune response to AAV during natural infection, sera from different age groups and various geographical regions were compared for AAV antibodies using an ELISA. The data show that the prevalence of antibodies to AAV is similar in Europe (Germany, France, and Switzerland), Brazil, and Japan, indicating worldwide infection. It was confirmed that infection takes place during childhood. However, declining seropositivity thereafter and a second increase of seropositivity after 30 years of age suggests reinfection or reactivation of latent virus in particular as the prevalence of IgM antibodies in adults is relatively high. Furthermore, pregnant women were found to be significantly more frequently seropositive than non-pregnant controls, hinting at a reactivation of persistent AAV (up to 80% of women carry AAV in genital tissue) in specific hormonal conditions, e.g., pregnancy. Cross-reaction of serum antibodies with the different AAV types (defined by complement fixation) was observed by ELISA and neutralization tests confirming earlier results. The results suggest an unstable AAV antibody response allowing lifelong reinfection or reactivation of persisting virus possibly due to partial immunotolerance after an infection in utero, at delivery or during early infancy.

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Subcellular compartmentalization of adeno-associated virus type 2 assembly

Cited by 181 publications

References 67 publications

Involvement of the nucleolus in replication of human viruses

Involvement of the nucleolus in replication of human viruses

The Role of the Adeno-Associated Virus Capsid in Gene Transfer

Update on the prevalence of serum antibodies (IgG and IgM) to adeno-associated virus (AAV)

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