Characterization of the POL3 gene product from Schizosaccharomyces pombe indicates inter-species conservation of the catalytic subunit of DNA polymerase delta

J Mol Biol. 1991 Nov 20;222(2):209-18. doi: 10.1016/0022-2836(91)90207-m.

Abstract

The Schizosaccharomyces pombe POL3 gene was isolated by sequence homology with a region of the Saccharomyces cerevisiae POL3 gene, the only gene sequenced to date encoding the catalytic subunit of eukaryotic DNA polymerase delta. The fission yeast POL3 gene contains a 52 base-pair (bp) intron and encodes a 3600 bp transcript the 5'-end of which is located 32 bp upstream from the initiation codon. The polypeptides predicted from budding and fission yeast POL3 genes share 52% of conserved amino acid residues and have a 60% identical central region. This structural conservation of the catalytic subunit of DNA polymerases delta is probably related to functional constraints. A portion of the most conserved region was used to raise antibodies against an S. pombe polymerase delta/beta-galactosidase fusion protein expressed in Escherichia coli. The purified antibodies recognized a 123,000 Da protein in S. pombe wild-type cell extracts and inhibited an aphidicolin-sensitive DNA polymerase activity that was distinct from DNA polymerase alpha. The antibodies also detected a 140,000 Da protein in extracts from different proliferating mammalian cells, indicating that the catalytic subunits of DNA polymerase delta are highly conserved between yeast and higher eukaryotes.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Blotting, Western
  • Chromosome Mapping
  • Cloning, Molecular
  • Cross Reactions
  • DNA Polymerase III
  • DNA-Directed DNA Polymerase / genetics*
  • DNA-Directed DNA Polymerase / immunology
  • Genes, Fungal*
  • Molecular Sequence Data
  • RNA, Fungal / genetics
  • RNA, Messenger / genetics
  • Restriction Mapping
  • Saccharomyces cerevisiae / enzymology
  • Schizosaccharomyces / enzymology
  • Schizosaccharomyces / genetics*
  • Sequence Alignment
  • Transcription, Genetic

Substances

  • RNA, Fungal
  • RNA, Messenger
  • DNA Polymerase III
  • DNA-Directed DNA Polymerase

Associated data

  • GENBANK/S58559
  • GENBANK/S58561
  • GENBANK/X56764
  • GENBANK/X56765
  • GENBANK/X56766
  • GENBANK/X56767
  • GENBANK/X56768
  • GENBANK/X56769
  • GENBANK/X59278
  • GENBANK/X62755