Half-calmodulin is sufficient for cell proliferation. Expressions of N- and C-terminal halves of calmodulin in the yeast Saccharomyces cerevisiae.
Calmodulin (CaM) has been shown to be an essential component for progression of nuclear division in the yeast Saccharomyces cerevisiae (Ohya, Y., and Anraku, Y. (1989) Curr. Genet. 15, 113-120). To define the functional domain of the molecule required for cell proliferation, we constructed plasmids expressing a series of N- and C-terminal halves of the CaM under the control of the galactose-inducible GAL1 promoter. These plasmids were introduced into a cmd1-disrupted yeast haploid strain, and the growth properties of the cells depending on the half-CaMs were examined. Plasmids expressing the N-terminal half (Ser1-Leu76) and the C-terminal half (Leu85-Cys147), which each maintain two complete EF-hand structures, complemented the growth defect of the cmd1 null mutation, whereas those expressing shorter regions of C- and N-terminal CaM did not. The half-CaMs that complemented the cmd1 null mutation were found to be approximately 6-fold overexpressed relative to expression of native CaM by the wild-type CMD1 gene. The levels of expression of the half CaMs with the true CMD1 promoter were not sufficient for complementation. These results demonstrate that half-CaMs (either the N- or the C-terminal) are capable of supporting growth of yeast cells when they are suitably overproduced. Cells depending solely on half-CaMs all showed a temperature-sensitive growth phenotype, suggesting that half-CaMs cannot carry out all the cellular functions of the complete CaM molecule.
Sun
G H
GH
Department of Biology, Faculty of Science, University of Tokyo, Japan.
Ohya
Y
Y
Anraku
Y
Y
D001483:Base Sequence; D002147:Calmodulin / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D002455:Cell Division; D002467:Cell Nucleus / Q000502:physiology / Q000648:ultrastructure; D004171:Diploidy; D005690:Galactose / Q000378:metabolism; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005947:Glucose / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015345:Oligonucleotide Probes; D010641:Phenotype; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics* / Q000254:growth & development; D013172:Spores, Fungal / Q000502:physiology
D002147:Calmodulin; D015345:Oligonucleotide Probes; D005947:Glucose; D005690:Galactose