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Volume 136, Issue 1

Calmodulin and calmodulin-binding proteins of , a basidiomycetous yeast Free

Abstract

Calmodulin (CaM)-interacting proteins of , a basidiomycetous yeast, were studied. CaM-binding proteins isolated from the soluble cell extract of vegetative cells and gamete cells (mating-pheromone-treated cells) by CaM-Sepharose affinity chromatography were analysed by sodium dodecylsulphate-polyacrylamide gel electrophoresis to compare the molecular species of CaM-binding proteins of the two cell types. The major CaM-binding proteins of the vegetative cells had molecular masses of 103 and 96 kDa. In the pheromone-treated cells, in addition to the bands seen in the vegetative cells, prominent bands of 40 and 38 kDa and minor bands of 92 and 60 kDa were present. The bands of CaM-binding proteins present in both cell types were more intense in the gamete cells. CaM-binding proteins of total cell extracts were also detected on acrylamide gels by a gel overlay method using I-CaM as the probe. The results obtained by the two procedures suggested that various CaM-binding proteins are present in the yeast cell, and the expression of many CaM-binding protein genes is altered by the mating pheromone. Ca- and CaM-dependent protein kinase activity was detected in the CaM-affinity-purified proteins. After incubation under conditions for protein phosphorylation by the kinase, the enzyme no longer required the activators, indicating that the enzyme activity is modulated by endogenous phosphorylation. A protein of 60 kDa was labelled by endogenous phosphorylation in a Ca/CaM-dependent manner, indicating that the labelled protein may be the Ca/CaM-dependent protein kinase or else be involved in its regulation.

  • Received:
  • Accepted:
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  • Published Online:
© Society for General Microbiology 1990
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1990-01-01
2024-05-01
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References

  1. Abe K., Kusaka I., Fukui S. 1975; Morphological change in the early stages of the mating process of Rhodosporidium toruloides. Journal of Bacteriology 122:710–718
     [Google Scholar]
  2. Baum P., Furlong S., Bayers B. 1986; Yeast gene required for spindle pole body duplication: homology of its product with Ca2+ binding proteins. Proceedings of the National Academy of Sciences of the United States of America 83:5512–5516
     [Google Scholar]
  3. Bolton A. E., Hunter W. M. 1973; The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent: application to the radioimmunoassay. Biochemical Journal 133:529–539
     [Google Scholar]
  4. Davis N. N., Urdea M. S., Masiarz F. R., Thorner J. 1986; Isolation of the yeast calmodulin gene: calmodulin is an essential protein. Cell 47:423–431
     [Google Scholar]
  5. Glenney J. R. Jr Weber K. 1983; Detection of calmodulinbinding polypeptides separated in SDS-polyacrylamide gels by a sensitive 125I calmodulin gel overlay assay. Methods in Enzymology 102:204–210
     [Google Scholar]
  6. Gopalakrishna R., Anderson W. B. 1982; Ca2+-induced hydrophobic site on calmodulin: application for purification of calmodulin by phenyl-sepharose affinity chromatography. Biochemical and Biophysical Research Communications 104:830–836
     [Google Scholar]
  7. Kamiya Y., Sakurai A., Tamura S., Takahashi N., Abe K., Tsuchiya E., Fukui S., Kitada C., Fujino M. 1979; Structure of rhodotorucine A, a peptidyl factor inducing mating tube formation in Rhodosporidium toruloides. Agricultural and Biological Chemistry 43:363–369
     [Google Scholar]
  8. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
     [Google Scholar]
  9. Lai Y., Nairn A. C., Greengard P. 1986; Autophosphorylation reversibly regulates the Ca2+/calmodulin-dependence of Ca2+/cal-modulin-dependent protein kinase II. Proceedings of the National Academy of Sciences of the United States of America 83:4253–4257
     [Google Scholar]
  10. Lou L. L., Lloyd S. J., Shulman H. 1986; Activation of the multifunctional Ca2+/calmodulin-dependent protein kinase by autophosphorylation: ATP modulated production of an autonomous enzyme. Proceedings of the National Academy of Sciences of the United States of America 83:9497–9501
     [Google Scholar]
  11. Miyakawa T., Nishihara M., Tsuchiya E., Fukui S. 1982; Role of the metabolism of the mating pheromone in sexual differentiation of the heterobasidiomycete Rhodosporidium toruloides. Journal of Bacteriology 151:1184–1194
     [Google Scholar]
  12. Miyakawa T., Tachikawa T., Jeong Y. K., Tsuchiya E., Fukui S. 1985; Transient increase of Ca2+ as a signal for mating pheromone induced differentiation in the heterobasidiomycetous yeast Rhodosporidium toruloides. Journal of Bacteriology 162:130–41306
     [Google Scholar]
  13. Miyakawa T., Tachikawa T., Akada R., Tsuchiya E., Fukui S. 1986; Involvement of Ca2+/calmodulin in the sexual differentiation induced by mating pheromone rhodotorucine λin Rhodosporidium toruloides. Journal of General Microbiology 132:1453–1457
     [Google Scholar]
  14. Miyakawa T., Oka Y., Tsuchiya E., Fukui S. 1989; Saccharomyces cerevisiae protein kinase dependent on Ca2+ and calmodulin. Journal of Bacteriology 171:1417–1422
     [Google Scholar]
  15. Ohya Y., Anraku Y. 1989; Functional expression of chicken calmodulin in yeast. Biochemical and Biophysical Research Communications 158:541–547
     [Google Scholar]
  16. Schworer C. M., Colbram R. J., Sorderling T. R. 1986; Reversible generation of a Ca2+(calmodulin)-dependent protein kinase II by an autophosphorylation mechanism. Journal of Biological Chemistry 261:8581–8584
     [Google Scholar]
  17. Takeda T., Yamamoto M. 1987; Analysis and in vivodisruption of the gene coding for calmodulin in Schizosaccharomyces pombe. Proceedings of the National Academy of Sciences of the United States of America 84:3580–3584
     [Google Scholar]
  18. Weiss B., Prozialeck W., Cimino M., Barnette M. S., Wallace T. L. 1980; Pharmacological regulation of calmodulin. Annals of the New York Academy of Sciences 356:319–345
     [Google Scholar]
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Calmodulin and calmodulin-binding proteins of Rhodosporidium toruloides, a basidiomycetous yeast
Microbiology 136, 131 (1990); https://doi.org/10.1099/00221287-136-1-131
/content/journal/micro/10.1099/00221287-136-1-131
/content/journal/micro/10.1099/00221287-136-1-131
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