Isolation and characterization of temperature-sensitive plc1 mutants of the yeast Saccharomyces cerevisiae.
The PLC1 gene of the yeast Saccharomyces cerevisiae has been discovered to encode a homolog of mammalian phosphoinositide-specific phospholipase C (PLC). Five temperature-sensitive plc1 mutants were isolated by in vitro mutagenesis with subsequent plasmid shuffling. All of the amino acid substitutions that caused a temperature-sensitive growth phenotype were located in the X or the Y region, both of which are conserved among PLC isoenzymes. The PLC activity of all products of mutant plc1 genes was dramatically lower than that of the wild-type product, indicating that PLC activity itself is important for cell growth. At the restrictive temperature, plc1 mutant cells ceased growth at random times during the cell cycle, a result that suggests that PLC1 is required at several or all stages of the cell cycle.
Yoko-o
T
T
Department of Plant Sciences, Graduate School of Science, University of Tokyo, Japan.
Kato
H
H
Matsui
Y
Y
Takenawa
T
T
Toh-e
A
A
D000595:Amino Acid Sequence; D005434:Flow Cytometry; D005819:Genetic Markers; D007527:Isoenzymes / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010727:Phosphoric Diester Hydrolases / Q000235:genetics*; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D013696:Temperature
D005819:Genetic Markers; D007527:Isoenzymes; D010727:Phosphoric Diester Hydrolases; D043265:Phosphatidylinositol Diacylglycerol-Lyase