COLD-SENSITIVE CELL-DIVISION-CYCLE MUTANTS OF YEAST: ISOLATION, PROPERTIES, AND PSEUDOREVERSION STUDIES

Don Moir ¹, Sue E. Stewart ¹, Barbara C. Osmond ¹, and David Botstein ¹

¹ Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

We isolated 18 independent recessive cold-sensitive cell-division-cycle (cdc) mutants of Saccharomyces cerevisiae, in nine complementation groups. Terminal phenotypes exhibited include medial nuclear division, cytokinesis, and a previously undescribed terminal phenotype consisting of cells with a single small bud and an undivided nucleus. Four of the cold-sensitive mutants proved to be alleles of CDC11, while the remaining mutants defined at least six new cell-division-cycle genes: CDC44, CDC45, CDC48, CDC49, CDC50 and CDC51.—Spontaneous revertants from cold-sensitivity of four of the medial nuclear division cs cdc mutants were screened for simultaneous acquisition of a temperature-sensitive phenotype. The temperature-sensitive revertants of four different cs cdc mutants carried single new mutations, called Sup/Ts to denote their dual phenotype: suppression of the cold-sensitivity and concomitant conditional lethality at 37°. Many of the Sup/Ts mutations exhibited a cell-division-cycle terminal phenotype at the high temperature, and they defined two new cdc genes (CDC46 and CDC47). Two cold-sensitive medial nuclear division cdc mutants representing two different cdc genes were suppressed by different Sup/Ts alleles of another gene which also bears a medial nuclear division function (CDC46). In addition, the cold-sensitive medial nuclear division cdc mutant csH80 was suppressed by a Sup/Ts mutation yielding an unbudded terminal phenotype with an undivided nucleus at the high temperature. This mutation was an allele of CDC32. These results suggest a pattern of interaction among cdc gene products and indicate that cdc gene proteins might act in the cell cycle as complex specific functional assemblies.

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				G. Caponigro, M. R. Abedi, A. P. Hurlburt, A. Maxfield, W. Judd, and A. Kamb Transdominant genetic analysis of a growth control pathway PNAS, June 23, 1998; 95(13): 7508 - 7513. [Abstract] [Full Text] [PDF]

				F. Madeo, J. Schlauer, H. Zischka, D. Mecke, and K.-U. Frohlich Tyrosine Phosphorylation Regulates Cell Cycle-dependent Nuclear Localization of Cdc48p Mol. Biol. Cell, January 1, 1998; 9(1): 131 - 141. [Abstract] [Full Text]

				F. Madeo, E. Frohlich, and K.-U. Frohlich A Yeast Mutant Showing Diagnostic Markers of Early and Late Apoptosis J. Cell Biol., November 3, 1997; 139(3): 729 - 734. [Abstract] [Full Text] [PDF]

				S. Donovan, J. Harwood, L. S. Drury, and J. F. X. Diffley Cdc6p-dependent loading of Mcm proteins onto pre-replicative chromatin in budding yeast PNAS, May 27, 1997; 94(11): 5611 - 5616. [Abstract] [Full Text] [PDF]

				C. F. J. Hardy, O. Dryga, S. Seematter, P. M. B. Pahl, and R. A. Sclafani mcm5/cdc46-bob1 bypasses the requirement for the S phase activator Cdc7p PNAS, April 1, 1997; 94(7): 3151 - 3155. [Abstract] [Full Text] [PDF]

				B. Stillman Cell Cycle Control of DNA Replication Science, December 6, 1996; 274(5293): 1659 - 1663. [Abstract] [Full Text]

				J F Diffley Once and only once upon a time: specifying and regulating origins of DNA replication in eukaryotic cells. Genes & Dev., November 15, 1996; 10(22): 2819 - 2830. [PDF]

				N Okishio, Y Adachi, and M Yanagida Fission yeast Nda1 and Nda4, MCM homologs required for DNA replication, are constitutive nuclear proteins J. Cell Sci., January 2, 1996; 109(2): 319 - 326. [Abstract] [PDF]

				H Yan, A M Merchant, and B K Tye Cell cycle-regulated nuclear localization of MCM2 and MCM3, which are required for the initiation of DNA synthesis at chromosomal replication origins in yeast. Genes & Dev., November 1, 1993; 7(11): 2149 - 2160. [Abstract] [PDF]

				B J Stevenson, N Rhodes, B Errede, and G F Sprague Constitutive mutants of the protein kinase STE11 activate the yeast pheromone response pathway in the absence of the G protein. Genes & Dev., July 1, 1992; 6(7): 1293 - 1304. [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]

				E J Strauss and C Guthrie A cold-sensitive mRNA splicing mutant is a member of the RNA helicase gene family. Genes & Dev., April 1, 1991; 5(4): 629 - 641. [Abstract] [PDF]

				K.M. Hennessy and D. Botstein Regulation of DNA Replication during the Yeast Cell Cycle Cold Spring Harb Symp Quant Biol, January 1, 1991; 56(0): 279 - 284. [Abstract] [PDF]

				K M Hennessy, C D Clark, and D Botstein Subcellular localization of yeast CDC46 varies with the cell cycle. Genes & Dev., December 1, 1990; 4(12b): 2252 - 2263. [Abstract] [PDF]

				U Vijayraghavan, M Company, and J Abelson Isolation and characterization of pre-mRNA splicing mutants of Saccharomyces cerevisiae. Genes & Dev., August 1, 1989; 3(8): 1206 - 1216. [Abstract] [PDF]

				M Gatti and B S Baker Genes controlling essential cell-cycle functions in Drosophila melanogaster. Genes & Dev., April 1, 1989; 3(4): 438 - 453. [Abstract] [PDF]

				A. Adams, D Botstein, and D. Drubin A yeast actin-binding protein is encoded by SAC6, a gene found by suppression of an actin mutation Science, January 13, 1989; 243(4888): 231 - 233. [Abstract] [PDF]

				K.L. Clark, N.G. Davis, D.K. Wiest, J.-J. Hwang-Shum, and G.F. Sprague Jr. Response of Yeast {alpha} Cells to a-Factor Pheromone: Topology of the Receptor and Identification of a Component of the Response Pathway Cold Spring Harb Symp Quant Biol, January 1, 1988; 53(0): 611 - 620. [Abstract] [PDF]

				T. W. Burke, J. G. Cook, M. Asano, and J. R. Nevins Replication Factors MCM2 and ORC1 Interact with the Histone Acetyltransferase HBO1 J. Biol. Chem., April 27, 2001; 276(18): 15397 - 15408. [Abstract] [Full Text] [PDF]

				J. R. Lamb, V. Fu, E. Wirtz, and J. D. Bangs Functional Analysis of the Trypanosomal AAA Protein TbVCP with trans-Dominant ATP Hydrolysis Mutants J. Biol. Chem., June 8, 2001; 276(24): 21512 - 21520. [Abstract] [Full Text] [PDF]

				D. F. Crawford and H. Piwnica-Worms The G2 DNA Damage Checkpoint Delays Expression of Genes Encoding Mitotic Regulators J. Biol. Chem., September 28, 2001; 276(40): 37166 - 37177. [Abstract] [Full Text] [PDF]

				M. G. Alexandrow, M. Ritzi, A. Pemov, and J. L. Hamlin A Potential Role for Mini-chromosome Maintenance (MCM) Proteins in Initiation at the Dihydrofolate Reductase Replication Origin J. Biol. Chem., January 18, 2002; 277(4): 2702 - 2708. [Abstract] [Full Text] [PDF]