Isolation and analysis of the fission yeast gene encoding polymerase delta accessory protein PCNA.
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.
Waseem
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NH
CRC Cell Transformation Research Group, University of Dundee, UK.
Labib
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K
Nurse
P
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Lane
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DP
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004261:DNA Replication / Q000235:genetics; D004271:DNA, Fungal; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics*; D010957:Plasmids; D018809:Proliferating Cell Nuclear Antigen; D016601:RNA-Binding Proteins; D051381:Rats; D012568:Schizosaccharomyces / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins; D017386:Sequence Homology, Amino Acid; D013172:Spores, Fungal
D004271:DNA, Fungal; D005656:Fungal Proteins; D009687:Nuclear Proteins; C078746:PCN1 protein, S pombe; D018809:Proliferating Cell Nuclear Antigen; D016601:RNA-Binding Proteins; D029702:Schizosaccharomyces pombe Proteins