Roles for ORC in M phase and S phase

Science

Volumn 279, Issue 5357, 1998, Pages 1733-1737

  Dillin, Andrew ^a   Rine, Jasper ^a  

^a UNIVERSITY OF CALIFORNIA   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CELL CYCLE PROTEIN; DNA; PROTEIN; PROTEIN KINASE;

ALLELE; ARTICLE; CELL CYCLE; CELL CYCLE S PHASE; CHROMOSOME; CONTROLLED STUDY; ENZYME ACTIVITY; MUTATION; NONHUMAN; PHENOTYPE; PRIORITY JOURNAL; PULSED FIELD GEL ELECTROPHORESIS; SACCHAROMYCES CEREVISIAE; TEMPERATURE;

CDC28 PROTEIN KINASE, S CEREVISIAE; CELL CYCLE PROTEINS; CELL NUCLEUS; CHROMOSOMES, FUNGAL; CROSSES, GENETIC; DNA REPLICATION; DNA, FUNGAL; DNA-BINDING PROTEINS; FUNGAL PROTEINS; GENES, FUNGAL; MITOSIS; MUTATION; ORIGIN RECOGNITION COMPLEX; PHENOTYPE; S PHASE; SACCHAROMYCES CEREVISIAE; SACCHAROMYCES CEREVISIAE PROTEINS; TEMPERATURE;

FUNGI; SACCHAROMYCES CEREVISIAE;

EID: 0032513357     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.279.5357.1733     Document Type: Article

References (36)

1. S. P. Bell and B. Stillman, Nature 357, 128 (1992).
2. C. A. Fox, S. Loo, A. Dillin, J. Rine, Genes Dev. 9, 911 (1995).
3. C. Liang, M. Weinreich, B. Stillman, Cell 81, 667 (1995).
4. Reviewed in A. Dutta and S. P. Bell, Annu. Rev. Cell Biol. 13, 293 (1997)
5. P. B. Carpenter, P. R. Mueller, W. G. Dunphy, Nature 379, 357 (1996).
6. A. Rowles et al., Cell 87, 287 (1996).
7. P. Romanowski, M. A. Madine, A. Rowles, J. J. Blow, R. A. Laskey, Curr. Biol. 6, 1416 (1996).
8. G. Landis, R. Kelley, A. C. Spradling, J. Tower, Proc. Natl. Acad. Sci. U.S.A. 94, 3888 (1997).
9. J. F. Diffley, J. H. Cocker, S. J. Dowell, A. Rowley, Cell 78, 303 (1994).
10. J. J. Blow and R. A. Laskey, Nature 332, 546 (1988).
11. C. Santocanale and J. F. Diffley, EMBO J. 15, 6671 (1996).
12. S. Loo et al., Mol. Biol. Cell 6, 741 (1995).
13. O. M. Aparacio, D. M. Weinstein, S. P. Bell, Cell 91 59 (1997).
14. G. T. Maine, P. Sinha, B. K. Tye, Genetics 106, 365 (1984).
15. Y. Kubota, S. Mimura, S. Nishimoto, H. Takisawa, H. Nojima, Cell 81, 601 (1995).
16. S. P. Bell, R. Kobayashi, B. Stillman, Science 262, 1844 (1993).
17. 8 cells/ml. Gels were run on a BRL CHEF apparatus in the absence of ethidium bromide at 175 V for 15 hours with a 70-s switch interval followed by an additional 10 hours with a 120-s switch interval at 2°C.
18. K. M. Hennessy, A. Lee, E. Chen, D. Botstein, Genes Dev. 5, 958 (1991).
19. T. A. Weinert, G. L. Kiser, L. H. Hartwell, ibid. 8, 652 (1994).
20. 600) of 0.1 and incubated at 37°C. Samples were tested every hour for 9 hours. Cell viability was calculated as the number of viable colonies formed after plating at 23°C at each time point divided by the number of colonies that formed after plating at 23°C at the 0 hour time point.
21. Supplementary material is available at www. sciencemag.org/feature/data/974048.shl.
22. A. Dillin and J. Rine, data not shown.
23. U. Surana et al., Cell 65, 145 (1991).
24. 8) were harvested and lysed for each hourly time point.
25. The orc5 ts alleles were created by in vitro mutagenesis of the cloned ORC5 gene by the polymerase chain reaction (PCR). PCR-mutagenized DNA pools were cotransformed into yeast cells with a gapped plasmid missing most of the ORC5 reading frame such that in vivo recombination would restore plasmid-borne copies of ORC5. In one screen, an orc5-1 strain (JRY4253) was transformed and the transformants were screened for their inability to complement the ts defect of orc5-1 (34). Putative new ts alleles were then recovered and transformed into an orc5 Δ strain. orc5-2 through orc5-12 were identified in this manner. The remaining alleles were identified by directly transforming the orc5Δ strain (JRY4154) with the mutagenic PCR reaction and the gapped plasmid in a plasmid-shuffle protocol. Transformants were screened for their ability to complement orc5Δ at 23°C, but not at 37°C.
26. 600 = 0.05 for each time point. All strains were made p°, hence lacking mitochondrial DNA, to avoid interference of mitochondrial DNA.
27. 600 = 0.05 at either 37° or 23°C. For PFGE, cells were similarly released from the M phase block and cells were harvested after 200 min at either 37° or 23°C.
28. S).
29. J. Conde and G. R. Fink, Proc. Natl. Acad. Sci. U.S.A. 73, 3651 (1976).
30. M. D. Rose, Annu. Rev. Cell. Dev. Biol. 12, 663 (1996).
31. Supplementary material is available at www. sciencemag.org/feature/data/974048.shl.
32. S. Piatti, T. Bohm, J. H. Cocker, J. F. Diffley, K. Nasmyth, Genes Dev. 10, 1516 (1996).
33. C. S. Stueland, D. J. Lew, M. J. Cismowski, S. I. Reed, Mol. Cell. Biol. 13, 3744 (1993).
34. A. Dillin and J. Rine, Genetics 147, 1053 (1997).
35. r p° kar1-1) (M. Rose), JRY5487 (JRY3009 orc5Δp° pRS316-ORC5), JRY5493 (JRY3009 ADE2 lys2Δ hmrΔ::URA3 orc5-1).
36. We thank members of the Rine lab, especially M. Neff, for insightful discussions; C. Beh for help with the karyogamy experiments; M. Botchan, S. Martin, and S. Okamura for comments on the manuscript; M. Rose and T. Weinert for strains; and B. Hyun (University of California, San Francisco) for help with FACScan analysis. Supported by an NIH predoctoral fellowship and a Genentech Distinguished Predoctoral Research Fellowship to A.D., a grant from the National Institutes of Health (GM-31105), and by a Mutagenesis Center grant from the National Institute of Environmental Health Sciences for core support (P30ESO1896-12).