Abstract
The p34cdc2 protein kinase plays a key role in the control of the mitotic cell cycle of fission yeast, being required for both entry into S-phase and for entry into mitosis in the mitotic cell cycle, as well as for the initiation of the second meiotic nuclear division. In recent years, structural and functional homologues of p34cdc2, as well as several of the proteins that interact with and regulate p34cdc2 function in fission yeast, have been identified in a wide range of higher eukaryotic cell types, suggesting that the control mechanisms uncovered in this simple eukaryote are likely to be well conserved across evolution. Here we describe the construction and characterisation of a fission yeast strain in which the endogenous p34cdc2 protein is entirely absent and is replaced by its human functional homologue p34CDC2, We have used this strain to analyse aspects of the function of the human p34CDC2 protein genetically. We show that the function of the human p34CDC2 protein in fission yeast cells is dependent upon the action of the protein tyrosine phosphatase p80cdc25 that it responds to altered levels of both the mitotic inhibitor p1072331 and the p34cdc2-binding protein p13suc1, and is lethal in combination with the mutant B-type cyclin p56cdc13-117. In addition, we demonstrate that the human p34CDC2 protein is proficient for fission yeast meiosis, and examine the behaviour of two mutant p34CDC2 proteins in fission yeast.
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References
Beach D, Nurse P (1981) High frequency transformation of the fission yeast Schizosaccharomyces pombe. Nature 290:140–142
Booher R, Beach D (1987) Interaction between cdc2 + and cdc13 + in the control of mitosis in fission yeast; dissociation of the G1l and G2 roles of the cdc2 + protein kinase. EMBO J 6:3441–3447
Carr AM, MacNeill SA, Hayles J, Nurse P (1989) Molecular cloning and sequence analysis of mutant alleles of the fission yeast cdc2 protein kinase gene: implications for cdc2 + protein structure and function. Mol Gen Genet 218:41–49
Clarke PR, Karsenti E (1991) Regulation of p34cdc2 protein kinase: new insights into protein phosphorylation and the cell cycle. J Cell Sci 100:409–414
Enoch T, Nurse P (1990) Mutation of fission yeast cell cycle control genes abolishes dependence of mitosis on DNA replication. Cell 60:665–673
Fantes P (1981) Isolation of cell size mutants of the fission yeast by a new selective method: characterisation of mutants and implications for division control mechanisms. J Bacteriol 146:746–754
Featherstone C, Russell P (1991) Fission yeast p107wee1 mitotic inhibitor is a protein tyrosine/serine kinase. Nature 349:808–811
Fleig UN, Nurse P (1991) Expression of a dominant negative allele of cdc2 prevents activation of the endogenous p34cdc2 kinase. Mol Gen Genet 226:432–440
Galaktionov K, Beach D (1991) Specific activation of cdc25 tyrosine phosphatases by B-type cyclins: evidence for multiple roles of mitotic cyclins. Cell 67:1181–1194
Gould KL, Moreno S, Owen DJ, Sazer S, Nurse P (1991) Phosphorylation at Thr167 is required for fission yeast p34cdc2 function. EMBO J 10:3297–3309
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
Hagan IM, Hayles J, Nurse P (1989) Cloning and sequencing of the cyclin-related cdc13 + gene and a cytological study of its role in fission yeast mitosis. J Cell Sci 91:587–595
Hanks SK, Quinn AM, Hunter T (1988) The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science 241:42–52
Hayles J, Aves S, Nurse P (1986) sucl is an essential gene involved in both the cell cycle and growth in fission yeast. EMBO J 5:3373–3379
Hindley J, Phear G (1984) Sequence of the cell division gene CDC2 from Schizosaccharomyces pombe: patterns of splicing and homology to protein kinases. Gene 31:129–134
Igarashi M, Nagata A, Jinno S, Suto K, Okayama H (1991) Weel + like gene in human cells. Nature 353:80–83
Lee M, Nurse P (1987) Complementation used to clone a human homologue of the fission yeast cell cycle control gene cdc2. Nature 327:31–35
Lundgren K, Walworth N, Booher R, Dembski M, Kirschner M, Beach D (1991) mik1 and weel cooperate in the inhibitory tyrosine phosphorylation of cdc2. Cell 64:1115–1122
MacNeill SA, Nurse P (1993) Mutational analysis of the fission yeast p34cdc2 protein kinase. Mol Gen Genet 236:415–426
MacNeill SA, Warbrick E, Fantes PA (1991) Controlling cell cycle progress in the fission yeast Schizosaccharomyces pombe. Curr Opin Genet Dev 1:307–312
Millar JBA, Russell P (1992) The cdc25 M-phase inducer: an unconventional protein phosphatase. Cell 68:407–410
Millar JBA, McGowan CH, Lenaers G, Jones R, Russell P (1991) p80cdc25 mitotic inducer is the tyrosine phosphatase that activates p34cdc2 in fission yeast. EMBO J 10:4301–4309
Moreno S, Hayles J, Nurse P (1989) Regulation of p34cdc2 protein kinase during mitosis. Cell 58:361–372
Moreno S, Klar A, Nurse P (1990) Molecular genetic analysis- of fission yeast Schizosaccharomyces pombe. Methods Enzymol 194:795
Murray AW, Kirschner MW (1989) Cyclin synthesis drives the early embryonic cell cycle. Nature 339:275–280
Murray AW, Solomon MI, Kirschner MW (1989) The role of cyclin synthesis and degradation in the control of maturation promoting factor activity. Nature 339:280–286
Nagata A, Igarashi M, Jinno S, Suto K, Okayama H (1991) An additional homolog of the fission yeast cdc25 + gene occurs in humans and is highly expressed in some cancer cells. New Biologist 3:959–968
Nasmyth KA, Nurse P (1981) Cell division cycle mutants altered in DNA replication and mitosis in the fission yeast Schizosaccharomyces pombe. Mol Gen Genet 182:119–124
Niwa O, Yanagida M (1988) Universal and essential role of MPF/cdc2 +. Nature 336:430
Nurse P (1990) Universal control mechanism regulating onset of M-phase. Nature 344:503–508
Nurse P, Thuriaux P, Nasmyth KA (1976) Genetic control of the cell division cycle in the fission yeast Schizosaccharomyces pombe. Mol Gen Genet 146:167–178
Richardson HE, Stueland CS, Thomas J, Russell P, Reed SI (1990) Human cDNAs encoding homologs of the small p34cdc2a/cdc2 associated protein of Saccharomyces cerevisiae and Schizosaccharomyces pombe. Genes Dev 4:1332–1344
Rothstein RJ (1983) One step gene disruption in yeast. Methods Enzymol 101:202–211
Russell P, Nurse P (1987) Negative regulation of mitosis by weel +, a gene encoding a protein kinase homologue. Cell 45:145–153
Sadhu K, Reed SI, Richardson H, Reed ST (1989) Putative mitotic inducer gene cdc25 in human cells is predominantly expressed in G2. Proc Natl Acad Sci USA 87:5139–5143
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Press, Laboratory Cold Spring Harbor, New York
Sazer S, Sherwood SW (1990) Mitochondrial growth and DNA synthesis occur in the absence of nuclear DNA replication in fission yeast. J Cell Sci 97:509–516
Simanis V, Nurse P (1986) The cell cycle control gene cdc2 + of fission yeast encodes a protein kinase potentially regulated by phosphorylation. Cell 45:261–268
Wittenberg C, Reed SI (1989) Conservation of the function and regulation with the CDC28/cdc2 protein kinase family: characterisation of human CDC2Hs protein kinase in Saccharomyces cerevisiae. Mol Cell Biol 9:4064–4068
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Communicated by B. Kilbey
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MacNeill, S.A., Nurse, P. Genetic analysis of human p34CDC2 function in fission yeast. Molec. Gen. Genet. 240, 315–322 (1993). https://doi.org/10.1007/BF00280381
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DOI: https://doi.org/10.1007/BF00280381