Adeno-associated virus Rep represses the human integration site promoter by two pathways that are similar to those required for the regulation of the viral p5 promoter.
Adeno-associated virus serotype 2 (AAV2) can efficiently replicate in cells that have been infected with helper viruses, such as adenovirus or herpesvirus. However, in the absence of helper virus infection, AAV2 establishes latency by integrating its genome site specifically into PPP1R12C, a gene located on chromosome 19. This integration target site falls into one of the most gene-dense regions of the human genome, thus inviting the question as to whether the virus has evolved mechanisms to control this complex transcriptional environment in order to facilitate integration, maintain an apparently innocuous latency, and/or establish conditions that are conducive to the rescue of the integrated viral genome. The viral replication (Rep) proteins control and direct every known aspect of the viral life cycle and have been shown to tightly control all AAV2 promoters. In addition, a number of heterologous promoters are repressed by the AAV2 Rep proteins. Here, we demonstrate that Rep proteins efficiently repress expression from the target site PPP1R12C promoter. We find evidence that this repression employs mechanisms similar to those described for Rep-mediated AAV2 p5 promoter regulation. Furthermore, we show that the repression of the cellular target site promoter is based on two distinct mechanisms, one relying on the presence of a functional Rep binding motif within the 5' untranslated region (UTR) of PPP1R12C, whereas the second pathway requires only an intact nucleoside triphosphate (NTP) binding site within the Rep proteins, indicating the possible reliance of this pathway on interactions of the Rep proteins with cellular proteins that mediate or regulate cellular transcription.


OBJECTIVE
The observation that repression of transcription from the adeno-associated virus serotype 2 (AAV2) p5 and integration target site promoters is mediated by shared mechanisms highlights the possible coevolution of virus and host and could lead to the identification of host factors that the virus exploits to navigate its life cycle.
Department of Infectious Diseases, King's College London School of Medicine, London, United Kingdom.
Nathalie
N
Dutheil
Department of Infectious Diseases, King's College London School of Medicine, London, United Kingdom.
Sarah C
SC
Smith
Department of Infectious Diseases, King's College London School of Medicine, London, United Kingdom.
Leticia
L
Agúndez
Department of Infectious Diseases, King's College London School of Medicine, London, United Kingdom.
Zoé I
ZI
Vincent-Mistiaen
Department of Infectious Diseases, King's College London School of Medicine, London, United Kingdom.
Carlos R
CR
Escalante
Department of Infectious Diseases, King's College London School of Medicine, London, United Kingdom.
R Michael
RM
Linden
Department of Infectious Diseases, King's College London School of Medicine, London, United Kingdom els.henckaerts@kcl.ac.uk.
Els
E
Henckaerts
D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism*; D000229:Dependovirus / Q000502:physiology*; D005786:Gene Expression Regulation; D054884:Host-Pathogen Interactions; D006801:Humans; D011401:Promoter Regions, Genetic; D054645:Protein Phosphatase 1 / Q000235:genetics*; D014764:Viral Proteins / Q000378:metabolism*; D016662:Virus Integration; D017735:Virus Latency
D004268:DNA-Binding Proteins; D014764:Viral Proteins; C068734:rep proteins, Adeno-associated virus 2; C540874:PPP1R12C protein, human; D054645:Protein Phosphatase 1