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Title: Interactions between the adeno-associated virus replication proteins and host cellular factors
Author: Smith, Sarah Claire
ISNI:       0000 0004 6058 129X
Awarding Body: King's College London
Current Institution: King's College London (University of London)
Date of Award: 2016
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Adeno-associated virus (AAV) is a small, ssDNA virus with a unique biphasic life cycle in which productive replication is dependent on both helper virus and host cellular factors. Unable to replicate autonomously, infection by AAV alone leads to the establishment of latency through either integration of the viral genome, or long-term episomal persistence. The complex AAV life cycle is orchestrated almost entirely by four isoforms of a single multifunctional viral nonstructural replication (Rep) protein. While it is known that Rep must interact with a multitude of host factors in order to complete the AAV life cycle, little is yet known about the nature of such interactions with respect to AAV gene regulation and the establishment of latency. Here, we used a screening method called BioID to identify new interaction partners for the Rep proteins, resulting in the identification of a number of interesting candidates involved in gene regulation, including the transcriptional corepressor KAP1. We show that KAP1 binds the latent AAV2 genome at the rep ORF, leading to trimethylation of AAV2-associated H3K9. We present evidence that helper viruses target KAP1 for degradation and demonstrate that antagonism of PP1α by Rep52/Rep78 further counteracts KAP1- mediated repression by enhancing nuclear levels of phosphorylated KAP1- S824, and that this interaction is essential for AAV2 transcription and replication. This work challenges the currently held model for AAV latency, and introduces not only a new viral mechanism for the counteraction of KAP1 repression, but also the notion that KAP1 targeting represents a conserved requirement for replication among DNA viruses.
Supervisor: Linden, Ralph Michael ; Henckaerts, Els Jeanny V. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available