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

NH WASEEM, K LABIB, P NURSE, D. P. Lane

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    Abstract

    Five monoclonal antibodies raised against rat PCNA cross-reacted with a similiar 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 DELTApcn1 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.

    Original languageEnglish
    Pages (from-to)5111-5120
    Number of pages10
    JournalEMBO Journal
    Volume11
    Issue number13
    Publication statusPublished - Dec 1992

    Keywords

    • CYCLIN
    • DNA REPLICATION
    • S-PHASE
    • SCHIZOSACCHAROMYCES-POMBE
    • MOLECULAR-CLONING
    • STRAND SYNTHESIS
    • FIELD GEL-ELECTROPHORESIS
    • CELL CYCLE
    • PCNA
    • REPLICATION INVITRO
    • EUKARYOTIC DNA-POLYMERASES
    • SACCHAROMYCES-CEREVISIAE
    • CELL NUCLEAR ANTIGEN

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