Abstract
Subcellular localization of type II cAMP-dependent protein kinase is determined by the interactions of the regulatory subunit (RII) with specific RII-anchoring proteins. By using truncated NH2-terminal RII beta fusion proteins expressed in Escherichia coli and the mitotic protein kinase p34cdc2 isolated from HeLa cells or starfish oocytes, we investigated the in vitro phosphorylation of RII beta by these kinases. The putative site for phosphorylation by the mitotic kinases is Thr-69 in the NH2-terminal domain of RII beta. This phosphorylation site matches the consensus sequence X(T/S)PX(K/R) for p34cdc2 recognition and belongs to a well-conserved sequence found in all RII beta sequences known to date. In contrast to phosphorylation by casein kinase II or the cAMP-dependent protein kinase catalytic subunit, phosphorylation of RII beta by mitotic kinases impaired its interaction with a well-known RII-anchoring protein, the neuronal microtubule-associated protein 2. The potential regulatory significance of the phosphorylation of this site on the interaction with microtubule-associated protein 2 and other RII-anchoring proteins and the physiological relevance of this cyclin B/p34cdc2 kinase-catalyzed modification of RII beta (or phosphorylation by other proline-directed protein kinases) are discussed.
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- Bailly E., Dorée M., Nurse P., Bornens M. p34cdc2 is located in both nucleus and cytoplasm; part is centrosomally associated at G2/M and enters vesicles at anaphase. EMBO J. 1989 Dec 20;8(13):3985–3995. doi: 10.1002/j.1460-2075.1989.tb08581.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bloom G. S., Luca F. C., Vallee R. B. Microtubule-associated protein 1B: identification of a major component of the neuronal cytoskeleton. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5404–5408. doi: 10.1073/pnas.82.16.5404. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Braun R. K., Vulliet P. R., Carbonaro-Hall D. A., Hall F. L. Phosphorylation of RII subunit and attenuation of cAMP-dependent protein kinase activity by proline-directed protein kinase. Arch Biochem Biophys. 1991 Aug 15;289(1):187–191. doi: 10.1016/0003-9861(91)90460-z. [DOI] [PubMed] [Google Scholar]
- Bregman D. B., Bhattacharyya N., Rubin C. S. High affinity binding protein for the regulatory subunit of cAMP-dependent protein kinase II-B. Cloning, characterization, and expression of cDNAs for rat brain P150. J Biol Chem. 1989 Mar 15;264(8):4648–4656. [PubMed] [Google Scholar]
- Carmichael D. F., Geahlen R. L., Allen S. M., Krebs E. G. Type II regulatory subunit of cAMP-dependent protein kinase. Phosphorylation by casein kinase II at a site that is also phosphorylated in vivo. J Biol Chem. 1982 Sep 10;257(17):10440–10445. [PubMed] [Google Scholar]
- Clegg C. H., Cadd G. G., McKnight G. S. Genetic characterization of a brain-specific form of the type I regulatory subunit of cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3703–3707. doi: 10.1073/pnas.85.11.3703. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Camilli P., Moretti M., Donini S. D., Walter U., Lohmann S. M. Heterogeneous distribution of the cAMP receptor protein RII in the nervous system: evidence for its intracellular accumulation on microtubules, microtubule-organizing centers, and in the area of the Golgi complex. J Cell Biol. 1986 Jul;103(1):189–203. doi: 10.1083/jcb.103.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fernandez A., Mery J., Vandromme M., Basset M., Cavadore J. C., Lamb N. J. Effective intracellular inhibition of the cAMP-dependent protein kinase by microinjection of a modified form of the specific inhibitor peptide PKi in living fibroblasts. Exp Cell Res. 1991 Aug;195(2):468–477. doi: 10.1016/0014-4827(91)90398-e. [DOI] [PubMed] [Google Scholar]
- Griffiths G., Hollinshead R., Hemmings B. A., Nigg E. A. Ultrastructural localization of the regulatory (RII) subunit of cyclic AMP-dependent protein kinase to subcellular compartments active in endocytosis and recycling of membrane receptors. J Cell Sci. 1990 Aug;96(Pt 4):691–703. doi: 10.1242/jcs.96.4.691. [DOI] [PubMed] [Google Scholar]
- Guilly M. N., Danon F., Brouet J. C., Bornens M., Courvalin J. C. Autoantibodies to nuclear lamin B in a patient with thrombopenia. Eur J Cell Biol. 1987 Apr;43(2):266–272. [PubMed] [Google Scholar]
- Hemmings B. A., Aitken A., Cohen P., Rymond M., Hofmann F. Phosphorylation of the type-II regulatory subunit of cyclic-AMP-dependent protein kinase by glycogen synthase kinase 3 and glycogen synthase kinase 5. Eur J Biochem. 1982 Oct;127(3):473–481. doi: 10.1111/j.1432-1033.1982.tb06896.x. [DOI] [PubMed] [Google Scholar]
- Hirsch A. H., Glantz S. B., Li Y., You Y., Rubin C. S. Cloning and expression of an intron-less gene for AKAP 75, an anchor protein for the regulatory subunit of cAMP-dependent protein kinase II beta. J Biol Chem. 1992 Feb 5;267(4):2131–2134. [PubMed] [Google Scholar]
- Jahnsen T., Hedin L., Kidd V. J., Beattie W. G., Lohmann S. M., Walter U., Durica J., Schulz T. Z., Schiltz E., Browner M. Molecular cloning, cDNA structure, and regulation of the regulatory subunit of type II cAMP-dependent protein kinase from rat ovarian granulosa cells. J Biol Chem. 1986 Sep 15;261(26):12352–12361. [PubMed] [Google Scholar]
- Keryer G., Rios R. M., Landmark B. F., Skalhegg B., Lohmann S. M., Bornens M. A high-affinity binding protein for the regulatory subunit of cAMP-dependent protein kinase II in the centrosome of human cells. Exp Cell Res. 1993 Feb;204(2):230–240. doi: 10.1006/excr.1993.1029. [DOI] [PubMed] [Google Scholar]
- Labbé J. C., Capony J. P., Caput D., Cavadore J. C., Derancourt J., Kaghad M., Lelias J. M., Picard A., Dorée M. MPF from starfish oocytes at first meiotic metaphase is a heterodimer containing one molecule of cdc2 and one molecule of cyclin B. EMBO J. 1989 Oct;8(10):3053–3058. doi: 10.1002/j.1460-2075.1989.tb08456.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Labbé J. C., Cavadore J. C., Dorée M. M phase-specific cdc2 kinase: preparation from starfish oocytes and properties. Methods Enzymol. 1991;200:291–301. doi: 10.1016/0076-6879(91)00147-o. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Leiser M., Rubin C. S., Erlichman J. Differential binding of the regulatory subunits (RII) of cAMP-dependent protein kinase II from bovine brain and muscle to RII-binding proteins. J Biol Chem. 1986 Feb 5;261(4):1904–1908. [PubMed] [Google Scholar]
- Levy F. O., Oyen O., Sandberg M., Taskén K., Eskild W., Hansson V., Jahnsen T. Molecular cloning, complementary deoxyribonucleic acid structure and predicted full-length amino acid sequence of the hormone-inducible regulatory subunit of 3'-5'-cyclic adenosine monophosphate-dependent protein kinase from human testis. Mol Endocrinol. 1988 Dec;2(12):1364–1373. doi: 10.1210/mend-2-12-1364. [DOI] [PubMed] [Google Scholar]
- Lohmann S. M., De Camilli P., Walter U. Type II cAMP-dependent protein kinase regulatory subunit-binding proteins. Methods Enzymol. 1988;159:183–193. doi: 10.1016/0076-6879(88)59019-5. [DOI] [PubMed] [Google Scholar]
- Lohmann S. M., DeCamilli P., Einig I., Walter U. High-affinity binding of the regulatory subunit (RII) of cAMP-dependent protein kinase to microtubule-associated and other cellular proteins. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6723–6727. doi: 10.1073/pnas.81.21.6723. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Luo Z., Shafit-Zagardo B., Erlichman J. Identification of the MAP2- and P75-binding domain in the regulatory subunit (RII beta) of type II cAMP-dependent protein kinase. Cloning and expression of the cDNA for bovine brain RII beta. J Biol Chem. 1990 Dec 15;265(35):21804–21810. [PubMed] [Google Scholar]
- Nigg E. A., Schäfer G., Hilz H., Eppenberger H. M. Cyclic-AMP-dependent protein kinase type II is associated with the Golgi complex and with centrosomes. Cell. 1985 Jul;41(3):1039–1051. doi: 10.1016/s0092-8674(85)80084-2. [DOI] [PubMed] [Google Scholar]
- Rangel-Aldao R., Rosen O. M. Dissociation and reassociation of the phosphorylated and nonphosphorylated forms of adenosine 3':5' -monophosphate-dependent protein kinase from bovine cardiac muscle. J Biol Chem. 1976 Jun 10;251(11):3375–3380. [PubMed] [Google Scholar]
- Rios R. M., Celati C., Lohmann S. M., Bornens M., Keryer G. Identification of a high affinity binding protein for the regulatory subunit RII beta of cAMP-dependent protein kinase in Golgi enriched membranes of human lymphoblasts. EMBO J. 1992 May;11(5):1723–1731. doi: 10.1002/j.1460-2075.1992.tb05224.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosen O. M., Erlichman J. Reversible autophosphorylation of a cyclic 3':5'-AMP-dependent protein kinase from bovine cardiac muscle. J Biol Chem. 1975 Oct 10;250(19):7788–7794. [PubMed] [Google Scholar]
- Rubino H. M., Dammerman M., Shafit-Zagardo B., Erlichman J. Localization and characterization of the binding site for the regulatory subunit of type II cAMP-dependent protein kinase on MAP2. Neuron. 1989 Nov;3(5):631–638. doi: 10.1016/0896-6273(89)90273-0. [DOI] [PubMed] [Google Scholar]
- Sandberg M., Taskén K., Oyen O., Hansson V., Jahnsen T. Molecular cloning, cDNA structure and deduced amino acid sequence for a type I regulatory subunit of cAMP-dependent protein kinase from human testis. Biochem Biophys Res Commun. 1987 Dec 31;149(3):939–945. doi: 10.1016/0006-291x(87)90499-2. [DOI] [PubMed] [Google Scholar]
- Sarkar D., Erlichman J., Rubin C. S. Identification of a calmodulin-binding protein that co-purifies with the regulatory subunit of brain protein kinase II. J Biol Chem. 1984 Aug 10;259(15):9840–9846. [PubMed] [Google Scholar]
- Scott J. D., Glaccum M. B., Zoller M. J., Uhler M. D., Helfman D. M., McKnight G. S., Krebs E. G. The molecular cloning of a type II regulatory subunit of the cAMP-dependent protein kinase from rat skeletal muscle and mouse brain. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5192–5196. doi: 10.1073/pnas.84.15.5192. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scott J. D., Stofko R. E., McDonald J. R., Comer J. D., Vitalis E. A., Mangili J. A. Type II regulatory subunit dimerization determines the subcellular localization of the cAMP-dependent protein kinase. J Biol Chem. 1990 Dec 15;265(35):21561–21566. [PubMed] [Google Scholar]
- Taylor S. S., Buechler J. A., Yonemoto W. cAMP-dependent protein kinase: framework for a diverse family of regulatory enzymes. Annu Rev Biochem. 1990;59:971–1005. doi: 10.1146/annurev.bi.59.070190.004543. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]