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J Biol Chem, Vol. 273, Issue 31, 19747-19755, July 31, 1998
From the Department of Biochemistry and Molecular Biophysics,
Washington University School of Medicine,
St. Louis, Missouri 63110
Yeast DNA polymerase
Characterization of the Two Small Subunits of Saccharomyces
cerevisiae DNA Polymerase
(Pol) has three
subunits of 125, 58, and 55 kDa. The gene for the 125-kDa catalytic
subunit (POL3) has been known for several years. Here we
describe the cloning of the genes for the 58- and 55-kDa subunits using
peptide sequence analysis and searching of the yeast genome data base.
The 58-kDa subunit, encoded by the POL31 gene, shows
23-28% sequence similarity to the 48-kDa subunit of human Pol and
to S. pombe Cdc1. POL31 is allelic to
HYS2 and SDP5. The 55-kDa subunit is encoded by the POL32 gene (ORF YJR043c in the yeast data base). Very
limited sequence similarity was observed between Pol32p and
Schizosaccharomyces pombe Cdc27, the functionally analogous
subunit in S. pombe Pol. The POL32 gene is
not essential, but a deletion mutant shows cold sensitivity for growth
and is sensitive to hydroxyurea and DNA damaging agents. In addition,
lethality was observed when the POL32 deletion mutation was
combined with conditional mutations in either the POL3 or
POL31 gene. Pol32
strains are weak
antimutators and are defective for damage-induced mutagenesis. The
POL32 gene product binds proliferating cell nuclear
antigen. A gel filtration analysis showed that Pol32p is a dimer in
solution. When POL31 and POL32 were
co-expressed in Escherichia coli, a tetrameric (Pol31p·Pol32p)2 species was detected by gel filtration,
indicating that the two subunits form a complex.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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