ALTERED FIDELITY OF MITOTIC CHROMOSOME TRANSMISSION IN CELL CYCLE MUTANTS OF S. CEREVISIAE

Leland H. Hartwell ¹ and David Smith ¹

¹ Department of Genetics SK-50, University of Washington, Seattle, Washington 98195

Thirteen of 14 temperature-sensitive mutants deficient in successive steps of mitotic chromosome transmission (cdc2, 4, 5, 6, 7, 8, 9, 13, 14, 15, 16, 17 and 20) from spindle pole body separation to a late stage of nuclear division exhibited a dramatic increase in the frequency of chromosome loss and/or mitotic recombination when they were grown at their maximum permissive temperatures. The increase in chromosome loss and/or recombination is likely to be due to the deficiency of functional gene product rather than to an aberrant function of the mutant gene product since the mutant alleles are, with one exception, recessive to the wild-type allele for this phenotype. The generality of this result suggests that a delay in almost any stage of chromosome replication or segregation leads to a decrease in the fidelity of mitotic chromosome transmission. In contrast, temperature-sensitive mutants defective in the control step of the cell cycle (cdc28), in cytokinesis (cdc3) or in protein synthesis (ils1) did not exhibit increased recombination or chromosome loss.—Based upon previous results with mutants and DNA-damaging agents in a variety of organisms, we suggest that the induction of mitotic recombination in certain mutants is due to the action of a repair pathway upon nicks or gaps left in the DNA. This interpretation is supported by the fact that the induced recombination is dependent upon the RAD52 gene product, an essential component in the recombinogenic DNA repair pathway. Gene products whose deficiency leads to induced recombination are, therefore, strong candidates for proteins that function in DNA metabolism. Among the mutants that induce recombination are those known to be defective in some aspect of DNA replication (cdc2, 6, 8, 9) as well as some mutants defective in the G2 (cdc13 and 17) and M (cdc5 and 14) phases of the mitotic cycle. We suggest that special aspects of DNA metabolism may be occurring in G2 and M in order to prepare the chromosomes for proper segregation.

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				C. I. Nugent, T. R. Hughes, N. F. Lue, and V. Lundblad Cdc13p: A Single-Strand Telomeric DNA-Binding Protein with a Dual Role in Yeast Telomere Maintenance Science, October 11, 1996; 274(5285): 249 - 252. [Abstract] [Full Text]

				A Bauer and R Kolling The SAC3 gene encodes a nuclear protein required for normal progression of mitosis J. Cell Sci., January 6, 1996; 109(6): 1575 - 1583. [Abstract] [PDF]

				R Rothstein and S Gangloff Hyper-recombination and Bloom's syndrome: microbes again provide clues about cancer. Genome Res., December 1, 1995; 5(5): 421 - 426. [PDF]

				C. H. Sommers, E. J. Miller, B. Dujon, S. Prakash, and L. Prakash Conditional Lethality of Null Mutations in RTH1 That Encodes the Yeast Counterpart of a Mammalian 5`- to 3`-Exonuclease Required for Lagging Strand DNA Synthesis in Reconstituted Systems J. Biol. Chem., March 3, 1995; 270(9): 4193 - 4196. [Abstract] [Full Text] [PDF]

				S. Cross, C. Sanchez, C. Morgan, M. Schimke, S Ramel, R. Idzerda, W. Raskind, and B. Reid A p53-dependent mouse spindle checkpoint Science, March 3, 1995; 267(5202): 1353 - 1356. [Abstract] [PDF]

				I. I. Ouspenski, U. W. Mueller, A. Matynia, S. Sazer, S. J. Elledge, and B. R. Brinkley Ran-binding Protein-1 Is an Essential Component of the Ran/RCC1 Molecular Switch System in Budding Yeast J. Biol. Chem., February 3, 1995; 270(5): 1975 - 1978. [Abstract] [Full Text] [PDF]

				T A Weinert, G L Kiser, and L H Hartwell Mitotic checkpoint genes in budding yeast and the dependence of mitosis on DNA replication and repair. Genes & Dev., March 15, 1994; 8(6): 652 - 665. [Abstract] [PDF]

				A Mac Auley, Z Werb, and P. Mirkes Characterization of the unusually rapid cell cycles during rat gastrulation Development, January 3, 1993; 117(3): 873 - 883. [Abstract] [PDF]

				B D Page and M Snyder CIK1: a developmentally regulated spindle pole body-associated protein important for microtubule functions in Saccharomyces cerevisiae. Genes & Dev., August 1, 1992; 6(8): 1414 - 1429. [Abstract] [PDF]

				K M Hennessy, A Lee, E Chen, and D Botstein A group of interacting yeast DNA replication genes. Genes & Dev., June 1, 1991; 5(6): 958 - 969. [Abstract] [PDF]

				T. Weinert and L. Hartwell The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae Science, July 15, 1988; 241(4863): 317 - 322. [Abstract] [PDF]

				S. T. Merino, W. J. Cummings, S. N. Acharya, and M. E. Zolan Replication-dependent early meiotic requirement for Spo11 and Rad50 PNAS, September 12, 2000; 97(19): 10477 - 10482. [Abstract] [Full Text] [PDF]