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J Virol, July 1998, p. 6228-6232, Vol. 72, No. 7
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Cloning and Functional Analysis of Kaposi's Sarcoma-Associated Herpesvirus DNA Polymerase and Its Processivity Factor

Kai Lin,1,2 Charlotte Y. Dai,3 and Robert P. Ricciardi1,2,*

Department of Microbiology, School of Dental Medicine,1 and Graduate Programs in Biochemistry and Molecular Biophysics2 and Cell and Molecular Biology,3 School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104

Received 29 December 1997/Accepted 25 March 1998

Kaposi's sarcoma-associated herpesvirus (KSHV), or human herpesvirus 8, is a newly identified virus with tumorigenic potential. Here, we cloned and expressed the DNA polymerase (Pol-8) of KSHV and its processivity factor (PF-8). Pol-8 bound specifically to PF-8 in vitro. Moreover, the DNA synthesis activity of Pol-8 was shown in vitro to be strongly dependent on PF-8. Addition of PF-8 to Pol-8 allowed efficient synthesis of fully extended DNA products corresponding to the full-length M13 template (7,249 nucleotides), whereas Pol-8 alone could incorporate only several nucleotides. The specificity of PF-8 and Pol-8 for each other was demonstrated by their inability to be functionally replaced by the DNA polymerases and processivity factors of herpes simplex virus 1 and human herpesvirus 6.

* Corresponding author. Mailing address: Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Levy Research Building, Rm. 221, 4010 Locust St., Philadelphia, PA 19104. Phone: (215) 898-3905. Fax: (215) 898-8385. E-mail: ricciardi{at}biochem.dental.upenn.edu.

J Virol, July 1998, p. 6228-6232, Vol. 72, No. 7
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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