Domains of p85cdc10 required for function of the fission yeast DSC-1 factor.
p85cdc10 is a component of the S.pombe DSC-1 complex, which is thought to mediate periodic transcription of genes in late G1. In order to understand the role of p85cdc10 in the function of this complex, we have analysed which domains of p85cdc10 are required for biological activity and the formation of a stable DSC-1 complex in vitro, both in cdc10 temperature sensitive and null backgrounds. No DSC-1 activity is found in the absence of p85cdc10 and the activity of the complex is reduced or absent in all cdc10ts mutants tested. Full biological activity and rescue of a cdc10::ura4+ null allele requires the N-terminal domain, the cdc10/SWI6 repeats and the helical C-terminal region. In the absence of p85cdc10, both the C-terminal and cdc10/SWI6 repeat domains are required for DSC-1 activity in vitro. In a cdc10ts background, rescue of DSC-1 activity and complementation of mutants, requires only expression of the C-terminal domain, though the presence of the cdc10/SWI6 motifs enhances its activity. The N-terminal domain, alone, or in combination with the cdc10/SWI6 motifs, does not have biological activity, and does not restore DSC-1 activity. We conclude that both the C-terminal domain of p85cdc10 is critical for formation of the DSC-1 complex and that the cdc10/SWI6 motifs also play a role, perhaps by stabilizing the complex. Our data also suggest that the S.pombe DSC-1 complex contains more than one molecule of p85cdc10.
Reymond
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Unité de Recherches sur le Cycle Cellulaire, Swiss Institute for Experimental Cancer Research (ISREC), Epalinges.
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D001483:Base Sequence; D018797:Cell Cycle Proteins; D004271:DNA, Fungal; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D020558:GTP Phosphohydrolases; D007700:Kinetics; D046911:Macromolecular Substances; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D029701:Saccharomyces cerevisiae Proteins; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins; D013696:Temperature; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic
D018797:Cell Cycle Proteins; D004271:DNA, Fungal; D005656:Fungal Proteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; C075111:SWI6 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors; C061178:cdc10 protein, S pombe; C512892:CDC10 protein, S cerevisiae; D020558:GTP Phosphohydrolases