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. 1982;1(9):1063–1068. doi: 10.1002/j.1460-2075.1982.tb01297.x

A cAMP-dependent protein kinase is present in differentiating Dictyostelium discoideum cells

Jde Gunzburg 1, M Veron 1,*
PMCID: PMC553163  PMID: 16453431

Abstract

We demonstrate the occurrence of a cAMP-dependent protein kinase in Dictyostelium discoideum cells at the terminal stage of differentiation. A cAMP-binding component was purified to homogeneity by affinity chromatography. This subunit inhibits the activity of purified catalytic subunit from beef heart protein kinase; the inhibition is reversed upon addition of cAMP. The protein is highly specific for cAMP and has a dissociation constant of 4 nM. The isolated regulatory subunit is a monomer of 39 K, with a sedimentation coefficient of 3.5S and a frictional coefficient of 1.24. The differences between this regulatory subunit and regulatory subunits of protein kinases from other sources are discussed.

Keywords: Dictyostelium discoideum, cAMP, protein kinase, affinity chromatography

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  1. Alton T. H., Lodish H. F. Developmental changes in messenger RNAs and protein synthesis in Dictyostelium discoideum. Dev Biol. 1977 Oct 1;60(1):180–206. doi: 10.1016/0012-1606(77)90118-x. [DOI] [PubMed] [Google Scholar]
  2. Alton T. H., Lodish H. F. Synthesis of developmentally regulated proteins in Dictyostelium discoideum which are dependent on continued cell-cell interaction. Dev Biol. 1977 Oct 1;60(1):207–216. doi: 10.1016/0012-1606(77)90119-1. [DOI] [PubMed] [Google Scholar]
  3. Blumberg D. D., Margolskee J. P., Barklis E., Chung S. N., Cohen N. S., Lodish H. F. Specific cell-cell contacts are essential for induction of gene expression during differentiation of Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1982 Jan;79(1):127–131. doi: 10.1073/pnas.79.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  5. Builder S. E., Beavo J. A., Krebs E. G. Stoichiometry of cAMP and 1,N6-etheno-cAMP binding to protein kinase. J Biol Chem. 1980 Mar 25;255(6):2350–2354. [PubMed] [Google Scholar]
  6. Cooper S., Chambers D. A., Scanlon S. Identification and characterization of the adenosine 3',5'-cyclic monophosphate binding proteins appearing during the development of Dictyostelium discoideum. Biochim Biophys Acta. 1980 May 7;629(2):235–242. doi: 10.1016/0304-4165(80)90097-5. [DOI] [PubMed] [Google Scholar]
  7. Corbin J. D., Sugden P. H., West L., Flockhart D. A., Lincoln T. M., McCarthy D. Studies on the properties and mode of action of the purified regulatory subunit of bovine heart adenosine 3':5'-monophosphate-dependent protein kinase. J Biol Chem. 1978 Jun 10;253(11):3997–4003. [PubMed] [Google Scholar]
  8. Darmon M., Brachet P., Da Silva L. H. Chemotactic signals induce cell differentiation in Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3163–3166. doi: 10.1073/pnas.72.8.3163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dills W. L., Jr, Beavo J. A., Bechtel P. J., Krebs E. G. Purification of rabbit skeletal muscle protein kinase regulatory subunit using cyclic adenosine-3':5'-monophosphate affinity chromatography. Biochem Biophys Res Commun. 1975 Jan 6;62(1):70–77. doi: 10.1016/s0006-291x(75)80406-2. [DOI] [PubMed] [Google Scholar]
  10. Erlichman J., Rubin C. S., Rosen O. M. Physical properties of a purified cyclic adenosine 3':5'-monophosphate-dependent protein kinase from bovine heart muscle. J Biol Chem. 1973 Nov 10;248(21):7607–7609. [PubMed] [Google Scholar]
  11. Gerisch G., Fromm H., Huesgen A., Wick U. Control of cell-contact sites by cyclic AMP pulses in differentiating Dictyostelium cells. Nature. 1975 Jun 12;255(5509):547–549. doi: 10.1038/255547a0. [DOI] [PubMed] [Google Scholar]
  12. Jacquet M., Part D., Felenbok B. Changes in the polyadenylated messenger RNA population during development of Dictyostelium discoideum. Dev Biol. 1981 Jan 15;81(1):155–166. doi: 10.1016/0012-1606(81)90358-4. [DOI] [PubMed] [Google Scholar]
  13. Kay R. R. cAMP and spore differentiation in Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1982 May;79(10):3228–3231. doi: 10.1073/pnas.79.10.3228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kemp B. E., Benjamini E., Krebs E. G. Synthetic hexapeptide substrates and inhibitors of 3':5'-cyclic AMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1038–1042. doi: 10.1073/pnas.73.4.1038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kemp B. E., Graves D. J., Benjamini E., Krebs E. G. Role of multiple basic residues in determining the substrate specificity of cyclic AMP-dependent protein kinase. J Biol Chem. 1977 Jul 25;252(14):4888–4894. [PubMed] [Google Scholar]
  16. Leichtling B. H., Majerfeld I. H., Coffman D. S., Rickenberg H. V. Identification of the regulatory subunit of a cAMP-dependent protein kinase in Dictyostelium discoideum. Biochem Biophys Res Commun. 1982 Apr 14;105(3):949–955. doi: 10.1016/0006-291x(82)91062-2. [DOI] [PubMed] [Google Scholar]
  17. Leichtling B. H., Spitz E., Rickenberg H. V. A cAMP-binding protein from Dictyostelium discoideum regulates mammalian protein kinase. Biochem Biophys Res Commun. 1981 May 29;100(2):515–522. doi: 10.1016/s0006-291x(81)80207-0. [DOI] [PubMed] [Google Scholar]
  18. Leichtling B. H., Tihon C., Spitz E., Rickenberg H. V. A cytoplasmic cAMP-binding protein in Dictyostelium discoideum. Dev Biol. 1981 Feb;82(1):150–157. doi: 10.1016/0012-1606(81)90436-x. [DOI] [PubMed] [Google Scholar]
  19. Miyamoto E., Petzold G. L., Kuo J. F., Greengard P. Dissociation and activation of adenosine 3',5'-monophosphate-dependent and guanosine 3',5'-monophosphate-dependent protein kinases by cyclic nucleotides and by substrate proteins. J Biol Chem. 1973 Jan 10;248(1):179–189. [PubMed] [Google Scholar]
  20. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  21. Muneyama K., Bauer R. J., Shuman D. A., Robins R. K., Simon L. N. Chemical synthesis and biological activity of 8-substituted adenosine 3',5'-cyclic monophosphate derivatives. Biochemistry. 1971 Jun 8;10(12):2390–2395. doi: 10.1021/bi00788a033. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Rahmsdorf H. J., Gerisch G. Specific binding proteins for cyclic AMP and cyclic GMP in Dictyostelium discoideum. Cell Differ. 1978 Oct;7(5):249–257. doi: 10.1016/0045-6039(78)90026-x. [DOI] [PubMed] [Google Scholar]
  24. Rannels S. R., Corbin J. D. Studies on the function of the two intrachain cAMP binding sites of protein kinase. J Biol Chem. 1981 Aug 10;256(15):7871–7876. [PubMed] [Google Scholar]
  25. Rannels S. R., Corbin J. D. Two different intrachain cAMP binding sites of cAMP-dependent protein kinases. J Biol Chem. 1980 Aug 10;255(15):7085–7088. [PubMed] [Google Scholar]
  26. Robertson A. D., Grutsch J. F. Aggregation in Dictyostelium discoideum. Cell. 1981 Jun;24(3):603–611. doi: 10.1016/0092-8674(81)90087-8. [DOI] [PubMed] [Google Scholar]
  27. Sampson J. Developmentally regulated cyclic AMP-dependent protein kinases in Dictyostelium discoideum. Cell. 1977 May;11(1):173–180. doi: 10.1016/0092-8674(77)90328-2. [DOI] [PubMed] [Google Scholar]
  28. Siegel L. M., Monty K. J. Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases. Biochim Biophys Acta. 1966 Feb 7;112(2):346–362. doi: 10.1016/0926-6585(66)90333-5. [DOI] [PubMed] [Google Scholar]
  29. Veron M., Patte J. C. Intracellular cAMP binding proteins in Dictyostelium discoidium: differences in properties of the proteins from vegetative and differentiated cells. Dev Biol. 1978 Apr;63(2):370–376. doi: 10.1016/0012-1606(78)90141-0. [DOI] [PubMed] [Google Scholar]
  30. Wallace L. J., Frazier W. A. Photoaffinity labeling of cyclic-AMP- and AMP-binding proteins differentiating Dictyostelium discoideum cells. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4250–4254. doi: 10.1073/pnas.76.9.4250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Watts D. J., Ashworth J. M. Growth of myxameobae of the cellular slime mould Dictyostelium discoideum in axenic culture. Biochem J. 1970 Sep;119(2):171–174. doi: 10.1042/bj1190171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weber W., Hilz H. Adenosine-3':5'-monophosphate-binding proteins from bovine kidney. Isolation by affinity chromatography and limited proteolysis of the regulatory subunit of protein kinase II. Eur J Biochem. 1978 Feb 1;83(1):215–225. doi: 10.1111/j.1432-1033.1978.tb12086.x. [DOI] [PubMed] [Google Scholar]
  33. Weber W., Vogel C. W., Hilz H. A new cAMP affinity matrix for the rapid purification of protein kinase regulatory subunits. FEBS Lett. 1979 Mar 1;99(1):62–66. doi: 10.1016/0014-5793(79)80249-5. [DOI] [PubMed] [Google Scholar]
  34. Williams J. G., Tsang A. S., Mahbubani H. A change in the rate of transcription of a eukaryotic gene in response to cyclic AMP. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7171–7175. doi: 10.1073/pnas.77.12.7171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zoller M. J., Kerlavage A. R., Taylor S. S. Structural comparisons of cAMP-dependent protein kinases I and II from porcine skeletal muscle. J Biol Chem. 1979 Apr 10;254(7):2408–2412. [PubMed] [Google Scholar]
  36. de Gunzburg J., Veron M. Intracellular adenosine 3',5'-phosphate binding proteins in Dictyostelium discoideum: partial purification and characterization in aggregation competent cells. Biochemistry. 1981 Aug 4;20(16):4547–4554. doi: 10.1021/bi00519a006. [DOI] [PubMed] [Google Scholar]

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