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Journal of Cell Science, Vol 108, Issue 9 3109-3118, Copyright © 1995 by Company of Biologists
JOURNAL ARTICLES |
G D'Urso, B Grallert and P Nurse
Cell Cycle Laboratory, Imperial Cancer Research Fund, London, UK.
Genetic analysis in the yeast Schizosaccharomyces pombe has shown that three genes cdc18, cut5, and cdt1, are essential for DNA synthesis and also for the checkpoint control that couples completion of DNA replication to the onset of mitosis. To test whether assembly of the replication initiation complex is an important element in the checkpoint control pathway we have investigated if DNA polymerase alpha (pol1), a component of the initiation complex, is essential for the S-phase checkpoint control. We show that germinating S. pombe spores disrupted for the pol1 gene enter mitosis despite defects in DNA synthesis. This is shown by monitoring septation index, DNA content, and by direct immunofluorescence of mitotic spindles using antibodies to alpha-tubulin. In addition we have isolated six temperature sensitive mutants in the pol1 gene that cause cell cycle arrest when grown at the nonpermissive temperature. Our experiments support a model in which DNA polymerase alpha, in addition to being part of the initiation complex, is required for a checkpoint signal that is activated as cells traverse START, and is essential to prevent mitosis until S phase has been completed. In contrast, proteins responsible for the elongation of DNA may not be necessary for this checkpoint signal.
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X. S. Ye, R. R. Fincher, A. Tang, K. K. McNeal, S. E. Gygax, A. N. Wexler, K. B. Ryan, S. W. James, and S. A. Osmani Proteolysis and Tyrosine Phosphorylation of p34cdc2/Cyclin B. THE ROLE OF MCM2 AND INITIATION OF DNA REPLICATION TO ALLOW TYROSINE PHOSPHORYLATION OF p34cdc2 J. Biol. Chem., December 26, 1997; 272(52): 33384 - 33393. [Abstract] [Full Text] [PDF]
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G. S. Brush, D. M. Morrow, P. Hieter, and T. J. Kelly The ATM homologue MEC1 is required for phosphorylation of replication protein A in yeast PNAS, December 24, 1996; 93(26): 15075 - 15080. [Abstract] [Full Text] [PDF]
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B Grallert and P Nurse The ORC1 homolog orp1 in fission yeast plays a key role in regulating onset of S phase. Genes & Dev., October 15, 1996; 10(20): 2644 - 2654. [Abstract] [PDF]
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