Isolation and characterization of fission yeast sns mutants defective at the mitosis-to-interphase transition.
pim1-d1ts was previously identified in a visual screen for fission yeast mutants unable to complete the mitosis-to-interphase transition. pim1+ encodes the guanine nucleotide exchange factor (GEF) for the spi1 GTPase. Perturbations of this GTPase system by either mutation or overproduction of its regulatory proteins cause cells to arrest with postmitotic condensed chromosomes, an unreplicated genome, and a wide medial septum. The septation phenotype of pim1-d1ts was used as the basis for a more extensive screen for this novel class of sns (septated, not in S-phase) mutants. Seventeen mutants representing 14 complementation groups were isolated. Three strains, sns-A3, sns-A5, and sns-A6, representing two different alleles, are mutated in the pim1+ gene. Of the 13 non-pim1ts sns complementation groups, 11 showed genetic interactions with the spi1 GTPase system. The genes mutated in 10 sns strains were synthetically lethal with pim1-d1, and six sns strains were hypersensitive to overexpression of one or more of the known components of the spil GTPase system. Epistasis analysis places the action of the genes mutated in nine of these strains downstream of pim1+ and the action of one gene upstream of pim1+. Three strains, sns-A2, sns-B1, and sns-B9, showed genetic interaction with the spil GTPase system in every test performed. sns-B1 and sns-B9 are likely to identify downstream targets, whereas sns-A2 is likely to identify upstream regulators of the spi1 GTPase system that are required for the mitosis-to-interphase transition.
Matynia
A
A
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston, Texas 77030, USA.
Mueller
U
U
Ong
N
N
Demeter
J
J
Granger
A L
AL
Hinata
K
K
Sazer
S
S
D000483:Alleles; D000595:Amino Acid Sequence; D004843:Epistasis, Genetic; D020558:GTP Phosphohydrolases / Q000235:genetics; D015870:Gene Expression; D005816:Genetic Complementation Test; D008040:Genetic Linkage; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics; D007399:Interphase / Q000235:genetics*; D008938:Mitosis / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D012568:Schizosaccharomyces / Q000166:cytology* / Q000235:genetics / Q000302:isolation & purification
D020662:Guanine Nucleotide Exchange Factors; C431743:p58pim1; D020558:GTP Phosphohydrolases