Isolation and analysis of the fission yeast gene encoding polymerase delta accessory protein PCNA

EMBO J. 1992 Dec;11(13):5111-20.

Abstract

Five monoclonal antibodies raised against rat PCNA cross-reacted with a similar protein in the fission yeast Schizosaccharomyces pombe. One of these was used to screen an S.pombe cDNA expression library. An incomplete cDNA was isolated and used to screen a genomic library, identifying a single gene, designated pcn1+ (proliferating cell nuclear antigen). The gene encodes a protein of 260 amino acids, with a deduced sequence 52% identical to human and rat PCNAs, which are 98.5% identical to each other. The budding yeast PCNA homologue POL30 is only 35% identical to the human and rat proteins. Pcn1 has a region near the C-terminus of particularly high homology to higher eukaryotic PCNA proteins. pcn1+ is essential for viability and delta pcn1 cells undergo aberrant DNA replication before cell cycle arrest. Overproduction of the protein leads to cell cycle delay in G2. Disruption of pcn1+ is complemented by the human PCNA gene, demonstrating that these genes are functional homologues.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Southern
  • Cloning, Molecular
  • DNA Replication / genetics
  • DNA, Fungal
  • Fungal Proteins / genetics*
  • Genes, Fungal*
  • Genetic Complementation Test
  • Humans
  • Molecular Sequence Data
  • Nuclear Proteins / genetics*
  • Plasmids
  • Proliferating Cell Nuclear Antigen
  • RNA-Binding Proteins
  • Rats
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces pombe Proteins*
  • Sequence Homology, Amino Acid
  • Spores, Fungal

Substances

  • DNA, Fungal
  • Fungal Proteins
  • Nuclear Proteins
  • PCN1 protein, S pombe
  • Proliferating Cell Nuclear Antigen
  • RNA-Binding Proteins
  • Schizosaccharomyces pombe Proteins

Associated data

  • GENBANK/D13199
  • GENBANK/D13200
  • GENBANK/D13201
  • GENBANK/D13202
  • GENBANK/D13203
  • GENBANK/M94783
  • GENBANK/M94784
  • GENBANK/M94785
  • GENBANK/X54857
  • GENBANK/X68109