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Genome Replication and Regulation of Viral Gene Expression | Spotlight

Involvement of SSRP1 in Latent Replication of Kaposi's Sarcoma-Associated Herpesvirus

Jianhong Hu, Eugene Liu, Rolf Renne
Jianhong Hu
Department of Molecular Genetics and Microbiology and UF Shands Cancer Center, University of Florida, Gainesville, Florida 32610
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Eugene Liu
Department of Molecular Genetics and Microbiology and UF Shands Cancer Center, University of Florida, Gainesville, Florida 32610
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Rolf Renne
Department of Molecular Genetics and Microbiology and UF Shands Cancer Center, University of Florida, Gainesville, Florida 32610
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  • For correspondence: rrenne@ufl.edu
DOI: 10.1128/JVI.00907-09
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ABSTRACT

Kaposi's sarcoma-associated herpesvirus (also named human herpesvirus 8) is a γ-herpesvirus that undergoes both lytic and latent infection. During latent infection, two viral elements are required: latency-associated nuclear antigen (LANA), which functions as an origin binding protein, and the latent origin, which resides within the terminal repeats (TRs) of the viral genome. Previously, we identified two cis-elements within the TRs which are required for latent DNA replication: two LANA binding sites (LBS1 and LBS2 [LBS1/2]) and a GC-rich replication element (RE) upstream of LBS1/2. To further characterize the RE, we constructed a 71-bp minimal replicon (MR) and performed a detailed mutational analysis. Our data indicate that the first 8 nucleotides within the RE are critical for replication. Moreover, both the position and the distance between the RE and LBS1/2 can affect origin replication activity, suggesting that the RE may function as a loading pad for cellular proteins involved in replication. Using biotinylated DNA fragments of wild-type or mutant MRs as probes, we identified 30 proteins that preferentially bind to the origin. Among these proteins, structure-specific recognition protein 1 (SSRP1), a subunit of the FACT complex, and telomeric repeat binding factor 2 (TRF2) formed complexes with LANA at the MR region. Furthermore, the small interfering RNA-based knockdown of SSRP1, but not the dominant-negative-based knockdown of TRF2, significantly decreased the efficiency of LANA-dependent DNA replication. These results indicate that SSRP1 is a novel cellular protein involved in LANA-dependent DNA replication.

  • Copyright © 2009 American Society for Microbiology
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Involvement of SSRP1 in Latent Replication of Kaposi's Sarcoma-Associated Herpesvirus
Jianhong Hu, Eugene Liu, Rolf Renne
Journal of Virology Oct 2009, 83 (21) 11051-11063; DOI: 10.1128/JVI.00907-09

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Involvement of SSRP1 in Latent Replication of Kaposi's Sarcoma-Associated Herpesvirus
Jianhong Hu, Eugene Liu, Rolf Renne
Journal of Virology Oct 2009, 83 (21) 11051-11063; DOI: 10.1128/JVI.00907-09
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KEYWORDS

Antigens, Viral
DNA replication
DNA, Viral
DNA-binding proteins
Herpesvirus 8, Human
High Mobility Group Proteins
Nuclear Proteins
Transcriptional Elongation Factors
Virus Latency
virus replication

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