The human cytomegalovirus (HCMV) UL44 gene product, polymerase accesso
ry protein, was cloned and expressed in Escherichia coli as a 53 000 M
W protein. The activity of HCMV DNA polymerase (Pol) alone and Pol/UL4
4 complex was evaluated in Pol assays designed specifically to elucida
te Pol/UL44 interactions. Addition of UL44 to HCMV Pol with primed, si
ngle-stranded DNA resulted in increased incorporation of nucleotides i
nto DNA, which was correlated with enhanced enzyme processivity. Sever
al deletion mutants which span the UL44 sequence were constructed and
examined for the ability to stimulate Pol activity and to bind double-
stranded DNA. The functional domains of UL44 protein were determined t
o reside within the N-terminal 309 amino acids of the wild type sequen
ce, since deletions within this region resulted in loss of DNA binding
and the ability to stimulate Pol. Deletion of C-terminal amino acids
310-433 had no effect on the ability of UL44 protein to increase the p
rocessivity of HCMV DNA Pol.