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 language | English |
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Pages (from-to) | 5111-5120 |
Number of pages | 10 |
Journal | EMBO Journal |
Volume | 11 |
Issue number | 13 |
Publication status | Published - 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