Human Kaposi's sarcoma herpesvirus processivity factor-8 functions as a dimer in DNA synthesis.

In the absence of other proteins, the DNA polymerase (Pol-8) of Kaposi's sarcoma herpesvirus incorporates only several nucleotides from a primer template. However, association with the Kaposi's sarcoma herpesvirus processivity factor (PF-8) enables Pol-8 to incorporate thousands of nucleotides. Unlike the well described sliding clamp processivity factors, eukaryotic proliferating cell nuclear antigen and Escherichia coli beta-subunit, PF-8 and other herpesvirus processivity factors do not require a clamp loader or ATP to bind to template DNA. To begin to understand the mechanism used by PF-8 to achieve processivity, we have now purified PF-8 and demonstrated that it is a dimer both in solution and on the DNA. Mutational analysis of the PF-8 protein (396R) indicates that residues between 277 and 304 as well as the N-terminal 21 amino acids are required for dimerization. The results further correlate PF-8 dimerization with binding to Pol-8 and stabilizing Pol-8 on primer template. Notably, although removal of only 26 residues from the C terminus of PF-8 does not affect its ability to form dimers on DNA or to bind Pol-8, only short DNA chains (<100 nucleotides) are synthesized. This indicates that full-length PF-8 is necessary to enable Pol-8 to incorporate thousands of nucleotides. Interestingly, cross-linking of the processivity factor UL44 of cytomegalovirus reveals that it is a dimer in solution also.