Identification of a fourth subunit of mammalian DNA polymerase δ

Abstract

A 12-kDa and two 25-kDa polypeptides were isolated with highly purified calf thymus DNA polymerase δ by conventional chromatography. A 16-mer peptide sequence was obtained from the 12-kDa polypeptide which matched a new open reading frame from a human EST (AA402118) encoding a hypothetical protein of unknown function. The protein was designated as p12. Human EST AA402118 was identified as the putative human homologue of Schizosaccharomyces pombe Cdm1 by a tBlastn search of the EST data base using S. pombe Cdm1. The open reading frame of human EST AA402118 encoded a polypeptide of 107 amino acids with a predicted molecular mass of 12.4 kDa, consistent with the experimental findings, p12 is 25% identical to S pombe Cdm1. Both of the 25-kDa polypeptide sequences matched the hypothetical KIAA0039 protein sequence, recently identified as the third subunit of pol δ. Western blotting of immunoaffinity purified calf thymus pol revealed the presence of p125, p50, p68 (the KIAA0039 product), and p12. With the identification of p12 mammalian pol δ can now be shown to consist of four subunits. These studies pave the way for more detailed analysis of the possible functions of the mammalian subunits of pol δ.