Deletion of SFI1, a novel suppressor of partial Ras-cAMP pathway deficiency in the yeast Saccharomyces cerevisiae, causes G(2) arrest.
When glucose is added to Saccharomyces cerevisiae cells grown into stationary phase or on non-fermentable carbon sources a rapid loss of heat stress resistance occurs. Mutants that retain high stress resistance after addition of glucose are called 'fil', for deficient in fermentation induced loss of stress resistance. Transformation of the fil1 mutant, which harbours a point mutation in adenylate cyclase, with a yeast gene library on a single copy plasmid resulted in transformants that were again stress-sensitive. One of the genes isolated in this way was a gene of previously unknown function. We have called it SFI1, for suppressor of fil1. SFI1 is an essential gene. Combination of Sfi1 and cAMP pathway mutations indicates that Sfi1 itself is not involved in the cAMP pathway. Conditional sfi1 mutants did not show enhanced heat resistance under the restrictive condition, whereas overexpression of SFI1 rendered cells heat-sensitive. Sfi1 may be a downstream target of the protein kinase A pathway, but its precise relationship with heat resistance remains unclear. Further analysis showed that Sfi1 is required for cell cycle progression, more specifically for progression through G(2)-M transition. Cells expressing SFI1 under the control of a galactose-inducible promoter arrest after addition of glucose as doublets of undivided mother and daughter cells. These doublets contain a single nucleus and lack mitotic spindles. Sfi1 shares homology with Xenopus laevis XCAP-C, a protein required for chromosome assembly. The conserved residues between these two proteins show a strong bias for charged amino acids. Hence, Sfi1 might be required for correct mitotic spindle assembly and its precise role might be in chromosome condensation. In conclusion, we have identified an essential function in the G(2)-M transition of the cell cycle for a yeast gene of previously unknown function.
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Laboratorium voor Moleculaire Celbiologie, Katholieke Universiteit Leuven, Leuven, Belgium.
Winderickx
J
J
Nauwelaers
D
D
Dumortier
F
F
De Doncker
A
A
Thevelein
J M
JM
Van Dijck
P
P
D001483:Base Sequence; D018797:Cell Cycle Proteins; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000378:metabolism*; D004271:DNA, Fungal / Q000032:analysis; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D016195:G2 Phase / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D016153:Genes, Suppressor; D005947:Glucose / Q000378:metabolism; D018869:Heat-Shock Response / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic; D014404:Tubulin / Q000032:analysis; D018631:ras Proteins / Q000378:metabolism*
D018797:Cell Cycle Proteins; D004271:DNA, Fungal; D011994:Recombinant Proteins; D012097:Repressor Proteins; C121820:SFI1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014404:Tubulin; D000242:Cyclic AMP; D018631:ras Proteins; D005947:Glucose