Identification, cloning and characterization of a novel 47 kDa murine PKA C subunit homologous to human and bovine Cβ2

BMC Biochemistry - Tập 7 Số 1 - 2006
Ane Funderud1, Heidi Hatledal Henanger1, Tilahun Tolesa Hafte1, Paul S. Amieux2, Sigurd Ørstavik1, Bjørn Steen Skålhegg1
1Department of Nutrition Research, Institute of Basic Medical Sciences, University of Oslo, PO Box 1046, Blindern, 0317, Oslo, Norway
2Department of Pharmacology, University of Washington School of Medicine, PO Box 357750, Seattle, WA, 98195-7750, USA

Tóm tắt

Abstract Background Two main genes encoding the catalytic subunits Cα and Cβ of cyclic AMP dependent protein kinase (PKA) have been identified in all vertebrates examined. The murine, bovine and human Cβ genes encode several splice variants, including the splice variant Cβ2. In mouse Cβ2 has a relative molecular mass of 38 kDa and is only expressed in the brain. In human and bovine Cβ2 has a relative molecular mass of 47 kDa and is mainly expressed in lymphoid tissues. Results We identified a novel 47 kDa splice variant encoded by the mouse Cβ gene that is highly expressed in lymphoid cells. Cloning, expression, and production of a sequence-specific antiserum and characterization of PKA catalytic subunit activities demonstrated the 47 kDa protein to be a catalytically active murine homologue of human and bovine Cβ2. Based on the present results and the existence of a human brain-specifically expressed Cβ splice variant designated Cβ4 that is identical to the former mouse Cβ2 splice variant, the mouse splice variant has now been renamed mouse Cβ4. Conclusion Murine lymphoid tissues express a protein that is a homologue of human and bovine Cβ2. The murine Cβ gene encodes the splice variants Cβ1, Cβ2, Cβ3 and Cβ4, as is the case with the human Cβ gene.

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Tài liệu tham khảo

Skalhegg BS, Tasken K: Specificity in the cAMP/PKA signaling pathway. Differential expression,regulation, and subcellular localization of subunits of PKA. Front Biosci. 2000, 5: D678-D693.

Rannels SR, Corbin JD: Two different intrachain cAMP binding sites of cAMP-dependent protein kinases. J Biol Chem. 1980, 255: 7085-7088.

Beebe SJ: The cAMP-dependent protein kinases and cAMP signal transduction. Semin Cancer Biol. 1994, 5: 285-294.

Amieux PS, McKnight GS: The essential role of RI alpha in the maintenance of regulated PKA activity. Ann N Y Acad Sci. 2002, 968: 75-95.

Zimmermann B, Chiorini JA, Ma Y, Kotin RM, Herberg FW: PrKX is a novel catalytic subunit of the cAMP-dependent protein kinase regulated by the regulatory subunit type I. J Biol Chem. 1999, 274: 5370-5378. 10.1074/jbc.274.9.5370.

Blaschke RJ, Monaghan AP, Bock D, Rappold GA: A novel murine PKA-related protein kinase involved in neuronal differentiation. Genomics. 2000, 64: 187-194. 10.1006/geno.2000.6116.

Reinton N, Haugen TB, Orstavik S, Skalhegg BS, Hansson V, Jahnsen T, et al.: The gene encoding the C gamma catalytic subunit of cAMP-dependent protein kinase is a transcribed retroposon. Genomics. 1998, 49: 290-297. 10.1006/geno.1998.5240.

Cummings DE, Edelhoff S, Disteche CM, McKnight GS: Cloning of a mouse protein kinase A catalytic subunit pseudogene and chromosomal mapping of C subunit isoforms. Mamm Genome. 1994, 5: 701-706. 10.1007/BF00426076.

Desseyn JL, Burton KA, McKnight GS: Expression of a nonmyristylated variant of the catalytic subunit of protein kinase A during male germ-cell development. Proc Natl Acad Sci U S A. 2000, 97: 6433-6438. 10.1073/pnas.97.12.6433.

Reinton N, Orstavik S, Haugen TB, Jahnsen T, Tasken K, Skalhegg BS: A novel isoform of human cyclic 3',5'-adenosine monophosphate-dependent protein kinase, c alpha-s, localizes to sperm midpiece. Biol Reprod. 2000, 63: 607-611. 10.1095/biolreprod63.2.607.

San Agustin JT, Leszyk JD, Nuwaysir LM, Witman GB: The catalytic subunit of the cAMP-dependent protein kinase of ovine sperm flagella has a unique amino-terminal sequence. J Biol Chem. 1998, 273: 24874-24883. 10.1074/jbc.273.38.24874.

Kvissel AK, Orstavik S, Oistad P, Rootwelt T, Jahnsen T, Skalhegg BS: Induction of Cbeta splice variants and formation of novel forms of protein kinase A type II holoenzymes during retinoic acid-induced differentiation of human NT2 cells. Cell Signal. 2004, 16: 577-587. 10.1016/j.cellsig.2003.08.014.

Showers MO, Maurer RA: A cloned bovine cDNA encodes an alternate form of the catalytic subunit of cAMP-dependent protein kinase. J Biol Chem. 1986, 261: 16288-16291.

Wiemann S, Kinzel V, Pyerin W: Isoform C beta 2, an unusual form of the bovine catalytic subunit of cAMP-dependent protein kinase. J Biol Chem. 1991, 266: 5140-5146.

Uhler MD, Chrivia JC, McKnight GS: Evidence for a second isoform of the catalytic subunit of cAMP-dependent protein kinase. J Biol Chem. 1986, 261: 15360-15363.

Guthrie CR, Skalhegg BS, McKnight GS: Two novel brain-specific splice variants of the murine Cbeta gene of cAMP-dependent protein kinase. J Biol Chem. 1997, 272: 29560-29565. 10.1074/jbc.272.47.29560.

Orstavik S, Reinton N, Frengen E, Langeland BT, Jahnsen T, Skalhegg BS: Identification of novel splice variants of the human catalytic subunit Cbeta of cAMP-dependent protein kinase. Eur J Biochem. 2001, 268: 5066-5073. 10.1046/j.0014-2956.2001.02429.x.

Orstavik S, Funderud A, Hafte TT, Eikvar S, Jahnsen T, Skalhegg BS: Identification and characterization of novel PKA holoenzymes in human T lymphocytes. FEBS J. 2005, 272: 1559-1567. 10.1111/j.1742-4658.2005.04568.x.

Howe DG, Wiley JC, McKnight GS: Molecular and behavioral effects of a null mutation in all PKA C beta isoforms. Mol Cell Neurosci. 2002, 20: 515-524. 10.1006/mcne.2002.1119.

Qi M, Zhuo M, Skalhegg BS, Brandon EP, Kandel ER, McKnight GS, Idzerda RL: Impaired hippocampal plasticity in mice lacking the Cbeta1 catalytic subunit of cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1996, 93: 1571-1576. 10.1073/pnas.93.4.1571.

Waterston RH, Lindblad-Toh K, Birney E, Rogers J, Abril JF, Agarwal P, et al.: Initial sequencing and comparative analysis of the mouse genome. Nature. 2002, 420: 520-562. 10.1038/nature01262.

Thullner S, Gesellchen F, Wiemann S, Pyerin W, Kinzel V, Bossemeyer D: The protein kinase A catalytic subunit Cbeta2: molecular characterization and distribution of the splice variant. Biochem J. 2000, 351: 123-132. 10.1042/0264-6021:3510123.

Beebe SJ, Oyen O, Sandberg M, Froysa A, Hansson V, Jahnsen T: Molecular cloning of a tissue-specific protein kinase (C gamma) from human testis--representing a third isoform for the catalytic subunit of cAMP-dependent protein kinase. Mol Endocrinol. 1990, 4: 465-475.

Skalhegg BS, Tasken K, Hansson V, Huitfeldt HS, Jahnsen T, Lea T: Location of cAMP-dependent protein kinase type I with the TCR-CD3 complex. Science. 1994, 263: 84-87.

Roskoski R: Assays of protein kinase. Methods Enzymol. 1983, 99: 3-6.

Skalhegg BS, Landmark BF, Doskeland SO, Hansson V, Lea T, Jahnsen T: Cyclic AMP-dependent protein kinase type I mediates the inhibitory effects of 3',5'-cyclic adenosine monophosphate on cell replication in human T lymphocytes. J Biol Chem. 1992, 267: 15707-15714.