Replication checkpoint enforced by kinases Cds1 and Chk1

Science

Volumn 280, Issue 5365, 1998, Pages 909-912

  Boddy, Michael N ^a   Furnari, Beth ^a   Mondesert, Odile ^a   Russell, Paul ^a  

^a SCRIPPS RESEARCH INSTITUTE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

HYDROXYUREA; PROTEIN KINASE;

ARTICLE; CELL DIVISION; ENZYME ACTIVATION; ENZYME PHOSPHORYLATION; NONHUMAN; PRIORITY JOURNAL; SCHIZOSACCHAROMYCES POMBE;

CDC2 PROTEIN KINASE; CDC25 PHOSPHATASE; CELL CYCLE; CELL CYCLE PROTEINS; DNA REPAIR; DNA REPLICATION; HYDROXYUREA; NUCLEAR PROTEINS; PHOSPHOPROTEIN PHOSPHATASE; PHOSPHORYLATION; PROTEIN KINASES; PROTEIN-SERINE-THREONINE KINASES; PROTEIN-TYROSINE KINASES; RECOMBINANT FUSION PROTEINS; S PHASE; SCHIZOSACCHAROMYCES; SCHIZOSACCHAROMYCES POMBE PROTEINS;

SCHIZOSACCHAROMYCES POMBE;

EID: 0032496322     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.280.5365.909     Document Type: Article

References (28)

1. L. H. Hartwell and T. A. Weinert, Science 246, 629 (1989); L. Hartwell, Cell 71, 543 (1992).
2. L. H. Hartwell and T. A. Weinert, Science 246, 629 (1989); L. Hartwell, Cell 71, 543 (1992).
3. S. J. Elledge, Science 274, 1664 (1996).
4. T. Enoch and P. Nurse, Cell 60, 665 (1990).
5. K. Lundgren et al., ibid. 64, 1111 (1991).
6. P. Jin, Y. Gu, D. O. Morgan, J. Cell Biol. 134, 963 (1996); A. Blasina, S. Paegle, C. H. McGowan, Mol. Biol. Cell 8, 1013 (1997).
7. P. Jin, Y. Gu, D. O. Morgan, J. Cell Biol. 134, 963 (1996); A. Blasina, S. Paegle, C. H. McGowan, Mol. Biol. Cell 8, 1013 (1997).
8. N. Rhind, B. Furnari, P. Russell, Genes Dev. 11, 504 (1997).
9. B. Furnari, N. Rhind, P. Russell, Science 277, 1495 (1997).
10. Y. Sanchez et al., ibid., p. 1497; C.-Y. Peng et al., ibid., p. 1501.
11. Y. Sanchez et al., ibid., p. 1497; C.-Y. Peng et al., ibid., p. 1501.
12. N. Walworth, S. Davey, D. Beach, Nature 363, 368 (1993);
13. F. al-Khodairy et al., Mol. Biol. Cell 5, 147 (1994).
14. N. Rhind and P. Russell, unpublished data.
15. M. N. Boddy and P. Russell, unpublished data.
16. R. Aligue, L. Wu, P. Russell, J. Biol. Chem. 272, 13320 (1997).
17. 70 truncation product were replicated with a precise fusion of GST and amino acids 1 to 72 of Wee1.
18. +. All strains were leu1-32 ura4-D18. All nmt1 promoter constructs were integrated at the leu1-32 locus. Growth media and general methods for S. pombe have been described [S. Moreno, A. Klar, P. Nurse, Methods Enzymol. 194, 795 (1991)]. Unless otherwise indicated, yeast cultures were grown in YES medium at 30°C. YES consists of glucose, yeast extract, and amino acid supplements. HU was used at a concentration of 12 mM. Purification of GST fusion proteins and hexahis-tagged proteins expressed in S. pombe, immunoblotting, and kinase assays were performed as described [K. Shiozaki and P. Russell, Nature 378, 739 (1995)].
19. +. All strains were leu1-32 ura4-D18. All nmt1 promoter constructs were integrated at the leu1-32 locus. Growth media and general methods for S. pombe have been described [S. Moreno, A. Klar, P. Nurse, Methods Enzymol. 194, 795 (1991)]. Unless otherwise indicated, yeast cultures were grown in YES medium at 30°C. YES consists of glucose, yeast extract, and amino acid supplements. HU was used at a concentration of 12 mM. Purification of GST fusion proteins and hexahis-tagged proteins expressed in S. pombe, immunoblotting, and kinase assays were performed as described [K. Shiozaki and P. Russell, Nature 378, 739 (1995)].
20. M. Fernandez-Sarabia, C. McInerny, P. Harris, C. Gordon, P. Fantes, Mol. Gen. Genet. 238, 241 (1993).
21. H. Murakami and H. Okayama, Nature 374, 817 (1995).
22. + construct (7).
23. The in vivo interaction between GST:Cds1 and Wee1 shown in Fig. 20 did not require HU treatment, presumably because GST:Cds1 was overexpressed.
24. 600 of 0.3 in YES medium and then diluted to 13,000 cells/ml in YES media containing HU (12 mM), followed by growth at 30°C for a further 8 hours. Samples (75 μl) were taken at regular intervals, plated onto YES plates, and grown for 4 days at 30°C to determine survival.
25. H. Lindsay et al., Genes Dev. 12, 382 (1998).
26. T. Enoch, A. M. Carr, P. Nurse, ibid. 6, 2035 (1993).
27. K. Shiozaki and P. Russell, Methods Enzymol. 283, 506 (1997).
28. We thank N. Rhind, J.-M. Brondello, C. McGowan, and K. Shiozaki for their advice and the Scripps Cell Cycle Groups for their support and encouragement. Funded by NIH.