Abstract
The protein kinase-encoding genes RCK1 and RCK2 from Saccharomyces cerevisiae have been identified as suppressors of Schizosaccharomyces pombe cell cycle checkpoint mutations. Upon expression of these genes, radiation resistance is partially restored in S. pombe mutants with checkpoint deficiencies, but not in mutants with DNA repair defects. Some checkpoint mutants are sensitive to the DNA synthesis inhibitor hydroxyurea, and this sensitivity is also suppressed by RCK1 and RCK2. The degree of suppression can be modulated by varying expression levels. Expression of RCK1 or RCK2 in S. pombe causes cell elongation and decelerated growth. Cells expressing these genes have a single nucleus and a 2n DNA content. We conclude that these genes act in S. pombe to prolong the G2 phase of the cell cycle.
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Al-Khodairy F, Carr AM (1992) DNA repair mutants defining G2 checkpoint pathways in Schizosaccharomyces pombe. EMBO J 11:1343–1350
Al-Khodairy F, Fotou E, Sheldrick KS, Griffiths DJF, Lehmann AR, Carr AM (1994) Identification and characterization of new elements involved in checkpoint and feedback controls in fission yeast. Mol Biol Cell 5:147–160
Amon A, Surana U, Muroff I, Nasmyth K (1992) Regulation of p34cdc28 tyrosine phosphorylation is not required for entry into mitosis in Saccharomyces cerevisiae. Nature 355:368–371
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) (1992) Current protocols in molecular biology. John Wiley and Sons, New York
Bach M-L (1987) Cloning and expression of the OMP decarboxylase gene ura4 from Schizosaccharomyces pombe. Curr Genet 12:527–534
Bankmann M, Prakash L, Prakash S (1992) Yeast RAD14 and human xeroderma pigmentosum group A DNA repair genes encode homologous proteins. Nature 355:555–558
Birkenbihl RP, Subramani S (1992) Cloning and characterization of rad21 an essential gene of Schizosaccharomyces pombe involved in DNA double-strand-break repair. Nucleic Acids Res 20:6605–6611
Booher R, Beach D (1987) Interaction between cdc13 + and cdc2 + in the control of mitosis in fission yeast; dissociation of the G1 and G2 roles of the cdc2+ protein kinase. EMBO J 6:3441–3447
Broach JR, Strathern JN, Hicks JB (1979) Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene. Gene 8:121–133
Dahlkvist A, Sunnerhagen P (1994) Two novel deduced serine/threonine protein kinases from Saccharomyces cerevisiae. Gene 139:27–33
Dasso M, Newport JW (1990) Completion of DNA replication is monitored by a feedback system that controls the initiation of mitosis in vitro: studies in Xenopus. Cell 61:811–823
Enoch T, Nurse P (1990) Mutation of fission yeast cell cycle control genes abolishes dependence of mitosis on DNA replication. Cell 60:665–673
Enoch T, Carr AM, Nurse P (1992) Fission yeast genes involved in coupling mitosis to completion of DNA replication. Genes Dev 6:2035–2046
Fikes JD, Becker DM, Winston F, Guarente L (1990) Striking conservation of TFIID in Schizosaccharomyces pombe and Saccharomyces cerevisiae. Nature 346:291–294
Grimm C, Kohli J, Murray J, Maundrell K (1988) Genetic engineering of Schizosaccharomyces pombe: a system for gene disruption and replacement using the ura4 gene as a selectable marker. Mol Gen Genet 215:81–86
Gulyas KD, Donahue TF (1992) SSL2, a suppressor of a stemloop mutation in the HIS4 leader encodes the yeast homolog of human ERCC-3. Cell 69:1031–1042
Gutz H, Heslot H, Leupold U, Loprieno N (1974) Schizosaccharomyces pombe. In: King RC (ed) Handbook of genetics, vol 1. Plenum, New York, pp 395–446
Hannan MA, Miller DR, Nasim A (1976) Changes in UV-inactivation kinetics and division delay in Schizosaccharomyces pombe strains during different growth phases. Radiat Res 68:469–479
Hartwell LH, Weinert TA (1989) Checkpoints: controls that ensure the order of cell cycle events. Science 246:629–634
Hindley J, Phear GA (1984) Sequence of the cell division gene CDC2 from Schizosaccharomyces pombe; patterns of splicing and homology to protein kinases. Gene 31:129–134
Hoffman CS, Winston F (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57:267–272
Inoue H, Nojima H, Okayama H (1990) High efficiency transformation of Escherichia coli with plasmids. Gene 96:23–28
Jimenez G, Yucel J, Rowley R, Subramani S (1992) The rad3 + gene of Schizosaccharomyces pombe is involved in multiple checkpoint functions and in DNA repair. Proc Natl Acad Sci USA 89:4952–4956
Kanter-Smoler G, Dahlkvist A, Sunnerhagen P (1994) Improved method for rapid transformation of intact Schizosaccharomyces pombe cells. Biotechniques 16:798–800
Kastan MB, Zhan QM, Eldeiry WS, Carrier F, Jacks T, Walsh WV, Plunkett BS, Vogelstein B, Fornace AJ (1992) A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia. Cell 71:587–597
Kuerbitz SJ, Plunkett BS, Walsh WV, Kastan MB (1992) Wild-type p53 is a cell cycle checkpoint determinant following irradiation. Proc Natl Acad Sci USA 89:7491–7495
Lee MG, Nurse P (1987) Complementation used to clone human homologue of the fission yeast cell cycle control gene cdc2. Nature 327:31–35
Lehmann AR, Carr AM, Watts FZ, Murray JM (1991) DNA repair in the fission yeast, Schizosaccharomyces pombe. Mutat Res 250:205–210
Lieberman HB, Hopkins KM, Laverty M, Chu HM (1992) Molecular cloning and analysis of Schizosaccharomyces pombe rad9, a gene involved in DNA repair and mutagenesis. Mol Gen Genet 232:367–376
Lorincz AT, Reed SI (1984) Primary structure homology between the product of yeast cell division control gene CDC28 and vertebrate oncogenes. Nature 307:183–185
Maundrell K (1990) nmt1 of fission yeast. J Biol Chem 265:10857–10864
Mitchinson JM (1974) Sequences pathways and timers in the cell cycle In: Padilla GM, Cameron IL, Zimmerman A (eds) Cell cycle controls. Academic Press, New York, pp 125–142
Moreno S, Klar A, Nurse P (1991) Molecular genetic analysis of the fission yeast Schizosaccharomyces pombe. Methods Enzymol 194:795–823
Murray JM, Carr AM, Lehmann AR, Watts FZ (1991) Cloning and characterization of the rad9 DNA repair gene from Schizosaccharomyces pombe. Nucleic Acids Res 19:3525–3531
Nasmyth KA, Tatchell K (1980) The structure of transposable yeast mating type loci. Cell 19:753–764
Picard D, Schena M, Yamamoto KR (1990) An inducible expression vector for both fission and budding yeast. Gene 86:257–261
Reed SI (1992) The role of p34 kinases in the G1 to S-phase transition. Annu Rev Cell Biol 8:529–661
Rowley R, Subramani S, Young PG (1992) Checkpoint controls in Schizosaccharomyces pombe - rad1. EMBO J 11:1335–1342
Russell P, Moreno S, Reed SI (1989) Conservation of mitotic controls in fission and budding yeasts. Cell 57:295–303
Schlegel R, Pardee AB (1986) Caffeine-induced uncoupling of mitosis from the completion of DNA replication in mammalian cells. Science 232:1264–1266
Seaton BL, Yucel J, Sunnerhagen P, Subramani S (1992) Isolation and characterization of the Schizosaccharomyces pombe rad3 gene involved in the DNA damage and DNA synthesis checkpoints. Gene 119:83–89
Sheldrick KS, Carr AM (1993) Feedback controls and G2 checkpoints: fission yeast as a model system. Bioessays 15:775–782
Sherr CJ (1993) Mammalian G1 cyclins. Cell 73:1059–1065
Shinohara A, Ogawa H, Matsuda Y, Ushio N, Ikeo K, Ogawa T (1993) Cloning of human mouse and fission yeast recombination genes homologous to RAD51 and recA. Nature Genetics 4:239–243
Sorger PK, Murray AW (1992) S-phase feedback control in budding yeast independent of tyrosine phosphorylation of p34cdc28. Nature 355:365–368
Stueland CS, Lew DJ, Cismowski MJ, Reed SI (1993) Full activation of p34cdc28 histone H1 kinase activity is unable to promote entry into mitosis in checkpoint-arrested cells of the yeast Saccharomyces cerevisiae. Mol Cell Biol 13:3744–3755
Sunnerhagen P, Seaton BL, Nasim A, Subramani S (1990) Cloning and analysis of a gene involved in DNA repair and recombination, the rad1 gene of Schizosaccharomyces pombe. Mol Cell Biol 10:3750–3760
Walworth N, Davey S, Beach D (1993) Fission yeast chk1 protein kinase links the rad checkpoint pathway to cdc2. Nature 363:368–371
Weinert TA, Hartwell LH (1988) The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae. Science 241:317–322
Weinert TA, Hartwell LH (1993) Cell cycle arrest of cdc mutants and specificity of the RAD9 checkpoint. Genetics 134:63–80
Zhou Z, Elledge SJ (1993) DUN1 encodes a protein kinase that controls the DNA damage response in yeast. Cell 75:1119–1127
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Dahlkvist, A., Kanter-Smoler, G. & Sunnerhagen, P. The RCK1 and RCK2 protein kinase genes from Saccharomyces cerevisiae suppress cell cycle checkpoint mutations in Schizosaccharomyces pombe . Molec. Gen. Genet. 246, 316–326 (1995). https://doi.org/10.1007/BF00288604
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DOI: https://doi.org/10.1007/BF00288604