DNA polymerase delta isolated from Schizosaccharomyces pombe contains five subunits.

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TitleDNA polymerase delta isolated from Schizosaccharomyces pombe contains five subunits.
Publication TypeJournal Article
Year of Publication1997
AuthorsZuo, S, Gibbs, E, Kelman, Z, Wang, TS, O'Donnell, M, MacNeill, SA, Hurwitz, J
JournalProc Natl Acad Sci U S A
Volume94
Issue21
Pagination11244-9
Date Published1997 Oct 14
ISSN0027-8424
KeywordsAmino Acid Sequence, Base Sequence, Blotting, Western, Chromatography, Affinity, Chromatography, Gel, Chromatography, Ion Exchange, DNA Polymerase III, DNA Replication, DNA, Fungal, DNA-Binding Proteins, Electrophoresis, Polyacrylamide Gel, Homeodomain Proteins, Humans, Kinetics, Macromolecular Substances, Molecular Sequence Data, Molecular Weight, Proliferating Cell Nuclear Antigen, Proto-Oncogene Proteins c-bcl-2, Replication Protein C, Repressor Proteins, Saccharomyces cerevisiae Proteins, Schizosaccharomyces
Abstract

DNA polymerase delta (pol delta) plays an essential role in DNA replication, repair, and recombination. We have purified pol delta from Schizosaccharomyces pombe more than 10(3)-fold and demonstrated that the polymerase activity of purified S. pombe pol delta is completely dependent on proliferating cell nuclear antigen and replication factor C. SDS/PAGE analysis of the purified fraction indicated that the pol delta complex consists of five subunits that migrate with apparent molecular masses of 125, 55, 54, 42, and 22 kDa. Western blot analysis indicated that the 125, 55, and 54 kDa proteins are the large catalytic subunit (Pol3), Cdc1, and Cdc27, respectively. The identity of the other two subunits, p42 and p22, was determined following proteolytic digestion and sequence analysis of the resulting peptides. The peptide sequences derived from the p22 subunit indicated that this subunit is identical to Cdm1, previously identified as a multicopy suppressor of the temperature-sensitive cdc1-P13 mutant, whereas peptide sequences derived from the p42 subunit were identical to a previously uncharacterized ORF located on S. pombe chromosome 1.

Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID9326594
PubMed Central IDPMC23429
Grant ListCA 54415 / CA / NCI NIH HHS / United States
GM 38559 / GM / NIGMS NIH HHS / United States
GM 38839 / GM / NIGMS NIH HHS / United States
/ / Wellcome Trust / United Kingdom