This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Hu, J.
Right arrow Articles by Renne, R.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hu, J.
Right arrow Articles by Renne, R.

 Previous Article  |  Next Article 

Journal of Virology, November 2009, p. 11051-11063, Vol. 83, No. 21
0022-538X/09/$08.00+0     doi:10.1128/JVI.00907-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

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

Jianhong Hu, Eugene Liu, and Rolf Renne*

Department of Molecular Genetics and Microbiology and UF Shands Cancer Center, University of Florida, Gainesville, Florida 32610

Received 6 May 2009/ Accepted 19 August 2009

Kaposi's sarcoma-associated herpesvirus (also named human herpesvirus 8) is a {gamma}-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.

* Corresponding author. Mailing address: University of Florida, 1376 Mowry Rd., Gainesville, FL 32610-0232. Phone: (352) 273-8204. Fax: (352) 273-8299. E-mail: rrenne{at}ufl.edu

{triangledown} Published ahead of print on 26 August 2009.

Journal of Virology, November 2009, p. 11051-11063, Vol. 83, No. 21
0022-538X/09/$08.00+0     doi:10.1128/JVI.00907-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»