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. 1987 Apr;7(4):1371–1377. doi: 10.1128/mcb.7.4.1371

Cloning and characterization of BCY1, a locus encoding a regulatory subunit of the cyclic AMP-dependent protein kinase in Saccharomyces cerevisiae.

T Toda, S Cameron, P Sass, M Zoller, J D Scott, B McMullen, M Hurwitz, E G Krebs, M Wigler
PMCID: PMC365223  PMID: 3037314

Abstract

We have cloned a gene (BCY1) from the yeast Saccharomyces cerevisiae that encodes a regulatory subunit of the cyclic AMP-dependent protein kinase. The encoded protein has a structural organization similar to that of the RI and RII regulatory subunits of the mammalian cyclic AMP-dependent protein kinase. Strains of S. cerevisiae with disrupted BCY1 genes do not display a cyclic AMP-dependent protein kinase in vitro, fail to grow on many carbon sources, and are exquisitely sensitive to heat shock and starvation.

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Selected References

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  1. Aiba H., Fujimoto S., Ozaki N. Molecular cloning and nucleotide sequencing of the gene for E. coli cAMP receptor protein. Nucleic Acids Res. 1982 Feb 25;10(4):1345–1361. doi: 10.1093/nar/10.4.1345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Bridgen P. J., Cross G. A., Bridgen J. N-terminal amino acid sequences of variant-specific surface antigens from Trypanosoma brucei. Nature. 1976 Oct 14;263(5578):613–614. doi: 10.1038/263613a0. [DOI] [PubMed] [Google Scholar]
  5. Builder S. E., Beavo J. A., Krebs E. G. The mechanism of activation of bovine skeletal muscle protein kinase by adenosine 3':5'-monophosphate. J Biol Chem. 1980 Apr 25;255(8):3514–3519. [PubMed] [Google Scholar]
  6. Choe J., Kolodrubetz D., Grunstein M. The two yeast histone H2A genes encode similar protein subtypes. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1484–1487. doi: 10.1073/pnas.79.5.1484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Constantinou A. I., Squinto S. P., Jungmann R. A. The phosphoform of the regulatory subunit RII of cyclic AMP-dependent protein kinase possesses intrinsic topoisomerase activity. Cell. 1985 Sep;42(2):429–437. doi: 10.1016/0092-8674(85)90100-x. [DOI] [PubMed] [Google Scholar]
  8. Cossart P., Gicquel-Sanzey B. Cloning and sequence of the crp gene of Escherichia coli K 12. Nucleic Acids Res. 1982 Feb 25;10(4):1363–1378. doi: 10.1093/nar/10.4.1363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dayhoff M. O., Barker W. C., Hunt L. T. Establishing homologies in protein sequences. Methods Enzymol. 1983;91:524–545. doi: 10.1016/s0076-6879(83)91049-2. [DOI] [PubMed] [Google Scholar]
  10. Gallwitz D., Sures I. Structure of a split yeast gene: complete nucleotide sequence of the actin gene in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 May;77(5):2546–2550. doi: 10.1073/pnas.77.5.2546. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hixson C. S., Krebs E. G. Characterization of a cyclic AMP-binding protein from bakers' yeast. Identification as a regulatory subunit of cyclic AMP-dependent protein kinase. J Biol Chem. 1980 Mar 10;255(5):2137–2145. [PubMed] [Google Scholar]
  12. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jurgensen S. R., Chock P. B., Taylor S., Vandenheede J. R., Merlevede W. Inhibition of the Mg(II).ATP-dependent phosphoprotein phosphatase by the regulatory subunit of cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7565–7569. doi: 10.1073/pnas.82.22.7565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kataoka T., Broek D., Wigler M. DNA sequence and characterization of the S. cerevisiae gene encoding adenylate cyclase. Cell. 1985 Dec;43(2 Pt 1):493–505. doi: 10.1016/0092-8674(85)90179-5. [DOI] [PubMed] [Google Scholar]
  15. Kataoka T., Powers S., Cameron S., Fasano O., Goldfarb M., Broach J., Wigler M. Functional homology of mammalian and yeast RAS genes. Cell. 1985 Jan;40(1):19–26. doi: 10.1016/0092-8674(85)90304-6. [DOI] [PubMed] [Google Scholar]
  16. Krebs E. G., Beavo J. A. Phosphorylation-dephosphorylation of enzymes. Annu Rev Biochem. 1979;48:923–959. doi: 10.1146/annurev.bi.48.070179.004423. [DOI] [PubMed] [Google Scholar]
  17. Matsumoto K., Uno I., Ishikawa T. Control of cell division in Saccharomyces cerevisiae mutants defective in adenylate cyclase and cAMP-dependent protein kinase. Exp Cell Res. 1983 Jun;146(1):151–161. doi: 10.1016/0014-4827(83)90333-6. [DOI] [PubMed] [Google Scholar]
  18. Matsumoto K., Uno I., Ishikawa T. Initiation of meiosis in yeast mutants defective in adenylate cyclase and cyclic AMP-dependent protein kinase. Cell. 1983 Feb;32(2):417–423. doi: 10.1016/0092-8674(83)90461-0. [DOI] [PubMed] [Google Scholar]
  19. Matsumoto K., Uno I., Oshima Y., Ishikawa T. Isolation and characterization of yeast mutants deficient in adenylate cyclase and cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2355–2359. doi: 10.1073/pnas.79.7.2355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Montgomery D. L., Leung D. W., Smith M., Shalit P., Faye G., Hall B. D. Isolation and sequence of the gene for iso-2-cytochrome c in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jan;77(1):541–545. doi: 10.1073/pnas.77.1.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nagamine Y., Reich E. Gene expression and cAMP. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4606–4610. doi: 10.1073/pnas.82.14.4606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Neff N. F., Thomas J. H., Grisafi P., Botstein D. Isolation of the beta-tubulin gene from yeast and demonstration of its essential function in vivo. Cell. 1983 May;33(1):211–219. doi: 10.1016/0092-8674(83)90350-1. [DOI] [PubMed] [Google Scholar]
  23. Ng R., Abelson J. Isolation and sequence of the gene for actin in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3912–3916. doi: 10.1073/pnas.77.7.3912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Potter R. L., Taylor S. S. Relationships between structural domains and function in the regulatory subunit of cAMP-dependent protein kinases I and II from porcine skeletal muscle. J Biol Chem. 1979 Apr 10;254(7):2413–2418. [PubMed] [Google Scholar]
  25. Potter R. L., Taylor S. S. The structural domains of cAMP-dependent protein kinase I. Characterization of two sites of proteolytic cleavage and homologies to cAMP-dependent protein kinase II. J Biol Chem. 1980 Oct 25;255(20):9706–9712. [PubMed] [Google Scholar]
  26. Rannels S. R., Corbin J. D. Characterization of small cAMP-binding fragments of cAMP-dependent protein kinases. J Biol Chem. 1979 Sep 10;254(17):8605–8610. [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Scott J. D., Fischer E. H., Demaille J. G., Krebs E. G. Identification of an inhibitory region of the heat-stable protein inhibitor of the cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4379–4383. doi: 10.1073/pnas.82.13.4379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Scott J. D., Fischer E. H., Takio K., Demaille J. G., Krebs E. G. Amino acid sequence of the heat-stable inhibitor of the cAMP-dependent protein kinase from rabbit skeletal muscle. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5732–5736. doi: 10.1073/pnas.82.17.5732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Smith M. M., Andrésson O. S. DNA sequences of yeast H3 and H4 histone genes from two non-allelic gene sets encode identical H3 and H4 proteins. J Mol Biol. 1983 Sep 25;169(3):663–690. doi: 10.1016/s0022-2836(83)80164-8. [DOI] [PubMed] [Google Scholar]
  31. Smith M., Leung D. W., Gillam S., Astell C. R., Montgomery D. L., Hall B. D. Sequence of the gene for iso-1-cytochrome c in Saccharomyces cerevisiae. Cell. 1979 Apr;16(4):753–761. doi: 10.1016/0092-8674(79)90091-6. [DOI] [PubMed] [Google Scholar]
  32. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  33. Takio K., Smith S. B., Krebs E. G., Walsh K. A., Titani K. Amino acid sequence of the regulatory subunit of bovine type II adenosine cyclic 3',5'-phosphate dependent protein kinase. Biochemistry. 1984 Aug 28;23(18):4200–4206. doi: 10.1021/bi00313a029. [DOI] [PubMed] [Google Scholar]
  34. Takio K., Walsh K. A., Neurath H., Smith S. B., Krebs E. G., Titani K. The amino acid sequence of a hinge region in the regulatory subunit of bovine cardiac muscle cyclic AMP-dependent protein kinase II. FEBS Lett. 1980 May 19;114(1):83–88. doi: 10.1016/0014-5793(80)80865-9. [DOI] [PubMed] [Google Scholar]
  35. Titani K., Sasagawa T., Ericsson L. H., Kumar S., Smith S. B., Krebs E. G., Walsh K. A. Amino acid sequence of the regulatory subunit of bovine type I adenosine cyclic 3',5'-phosphate dependent protein kinase. Biochemistry. 1984 Aug 28;23(18):4193–4199. doi: 10.1021/bi00313a028. [DOI] [PubMed] [Google Scholar]
  36. Toda T., Uno I., Ishikawa T., Powers S., Kataoka T., Broek D., Cameron S., Broach J., Matsumoto K., Wigler M. In yeast, RAS proteins are controlling elements of adenylate cyclase. Cell. 1985 Jan;40(1):27–36. doi: 10.1016/0092-8674(85)90305-8. [DOI] [PubMed] [Google Scholar]
  37. Uno I., Matsumoto K., Ishikawa T. Characterization of cyclic AMP-requiring yeast mutants altered in the regulatory subunit of protein kinase. J Biol Chem. 1982 Dec 10;257(23):14110–14115. [PubMed] [Google Scholar]
  38. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  39. Wallis J. W., Hereford L., Grunstein M. Histone H2B genes of yeast encode two different proteins. Cell. 1980 Dec;22(3):799–805. doi: 10.1016/0092-8674(80)90556-5. [DOI] [PubMed] [Google Scholar]
  40. Waterman M., Murdoch G. H., Evans R. M., Rosenfeld M. G. Cyclic AMP regulation of eukaryotic gene transcription by two discrete molecular mechanisms. Science. 1985 Jul 19;229(4710):267–269. doi: 10.1126/science.2990047. [DOI] [PubMed] [Google Scholar]
  41. Weber I. T., Takio K., Titani K., Steitz T. A. The cAMP-binding domains of the regulatory subunit of cAMP-dependent protein kinase and the catabolite gene activator protein are homologous. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7679–7683. doi: 10.1073/pnas.79.24.7679. [DOI] [PMC free article] [PubMed] [Google Scholar]

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