Inhibitors of protein kinases affecting cAMP-dependent proteolysis of GATA-6


We screened 95 kinase inhibitors whether they affect cAMP-dependent proteolysis of GATA-6 or not. Among them 7 inhibitors inhibited the proteolysis at the concentration range of μM around their IC50. They are inhibitors for protein kinase A (H-89 and 4- cyano-3-methylisoquinoline), c-Jun N-terminal kinase (SP600125), phosphatidylinositol 3-kinase (Wort- mannin and LY-294002), casein kinase II (TBB) and cyclin dependent kinase (Cdk1/2 inhibitor III). It is of interest how these kinases play roles in the degradation process of GATA-6 since this transcription factor is essential for development and tissue-specific gene expression of mammals. Inhibitors identified in this study would be helpful to study molecular mechanisms of phenomena in which GATA-6 participates.

Share and Cite:

Ushijima, H. and Maeda, M. (2012) Inhibitors of protein kinases affecting cAMP-dependent proteolysis of GATA-6. Advances in Biological Chemistry, 2, 411-415. doi: 10.4236/abc.2012.24051.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Tamura, S., Wang, X.H., Maeda, M., et al. (1993) Gastric DNA-binding proteins recognize upstream sequence motifs of parietal cell-specific genes. Proceedings of the National Academy of Sciences, 90, 10876-10880. doi:10.1073/pnas.90.22.10876
[2] Maeda, M., Kubo, K., Nishi, T., et al. (1996) Roles of gastric GATA DNA-binding proteins. The Journal of Experimental Biology, 199, 513-520.
[3] Maeda, M., Ohashi, K. and Ohashi-Kobayashi, A. (2005) Further extension of mammalian GATA-6. Development, Growth & Differentiation, 47, 591-600. doi:10.1111/j.1440-169X.2005.00837.x
[4] Nakagawa, R., Sato, R., Futai, M., et al. (1997) Gastric GATA-6 DNA-binding protein: Proteolysis induced by cAMP. FEBS Letters, 408, 301-305. doi:10.1016/S0014-5793(97)00443-2
[5] Ishida, A., Iijima, R., Kobayashi, A., et al. (2005) Characterization of cAMP-dependent proteolysis of GATA-6. Biochemical and Biophysical Research Communications, 332, 976-981. doi:10.1016/j.bbrc.2005.05.042
[6] Tsuge, T., Uetani, K., Sato, R., et al. (2008) Cyclic AMP-dependent proteolysis of GATA-6 expressed on the intracellular membrane. Cell Biology International, 32, 298-303. doi:10.1016/j.cellbi.2007.10.005
[7] Han, I. and Kudlow, J.E. (1997) Reduced O glycosylation of Sp1 is associated with increased proteasome susceptibility. Molecular and Cellular Biology, 17, 2550-2558.
[8] Su, K., Roos, M.D., Yang, X., et al. (1999) An N-terminal region of Sp1 targets its proteasome-dependent degradation in vitro. The Journal of Biological Chemistry, 274, 15194-15202. doi:10.1074/jbc.274.21.15194
[9] Maeda, M., Ishida, A., Ni, L., et al. (2005) Isolation of CHO-K1 clones defective in cAMP-dependent proteolysis, as determined by the stability of exogenously expressed GATA-6. Biochemical and Biophysical Research Communications, 329, 140-146. doi:10.1016/j.bbrc.2005.01.118
[10] Robinson-White, A. and Stratakis, C.A. (2002) Protein kinase A signaling: “cross-talk” with other pathways in endocrine cells. Annals of the New York Academy of Sciences, 968, 256-270. doi:10.1111/j.1749-6632.2002.tb04340.x
[11] SCADS inhibitor kit (2012).
[12] Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Bio- chemistry, 72, 248-254. doi:10.1016/0003-2697(76)90527-3
[13] Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685. doi:10.1038/227680a0
[14] Towbin, H., Staehelin, T. and Gordon, J. (1979) Electro-phoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proceedings of the National Academy of Sciences, 76, 4350-4354. doi:10.1073/pnas.76.9.4350
[15] Chijiwa, T., Mishima, A., Hagiwara, M., et al. (1990) Inhibition of forskolin-induced neurite outgrowth and protein phosphorylation by a newly synthesized selective inhibitor of cyclic AMP-dependent protein kinase, N-[2- (p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonam de (H-89), of PC12D pheochromocytoma cells. The Journal of Biological Chemistry, 265, 5267-5272.
[16] Lu, Z.X., Quazi, N.H., Deady, L.W., et al. (1996) Selective inhibition of cyclic AMP-dependent protein kinase by isoquinoline derivatives. Biological chemistry Hoppe- Seyler, 377, 373-384. doi:10.1515/bchm3.1996.377.6.373
[17] Lin, R., Connolly, P.J., Huang, S., et al. (2005) 1-Acyl- 1H-[1,2,4]triazole-3,5-diamine analogues as novel and potent anticancer cyclin-dependent kinase inhibitors: Synthesis and evaluation of biological activities. Journal of Medicinal Chemistry, 48, 4208-4211. doi:10.1021/jm050267e
[18] Szyszka, R., Grankowski, N., Felczak, K., et al. (1995) Halogenated benzimidazoles and benzotriazoles as selective inhibitors of protein kinases CK I and CK II from Saccharomyces cerevisiae and other sources. Biochemical and Biophysical Research Communications, 208, 418- 424. doi:10.1006/bbrc.1995.1354
[19] Bennett, B.L., Sasaki, D.T., Murray, B.W., et al. (2001) SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proceedings of the National Academy of Sciences, 98, 13681-13686. doi:10.1073/pnas.251194298
[20] Vlahos, C.J., Matter, W.F., Hui, K.Y., et al. (1994) A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4- morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). The Journal of Biological Chemistry, 269, 5241-5248.
[21] Powis, G., Bonjouklian, R., Berggren, M.M., et al. (1994) Wortmannin, a potent and selective inhibitor of phosphatidylinositol-3-kinase. Cancer Research, 54, 2419- 2423.
[22] Lee, M.G. and Nurse, P. (1987) Complementation used to clone a human homologue of the fission yeast cell cycle control gene cdc2. Nature, 327, 31-35. doi:10.1038/327031a0
[23] Tsai, L.H., Harlow, E. and Meyerson, M. (1991) Isolation of the human cdk2 gene that encodes the cyclin A- and adenovirus E1A-associated p33 kinase. Nature, 353, 174- 177. doi:10.1038/353174a0
[24] Allende, J.E. and Allende, C.C. (1995) Protein kinases. 4. Protein kinase CK2: An enzyme with multiple substrates and a puzzling regulation. FASEB Journal, 9, 313-323.
[25] Derijard, B., Hibi, M., Wu, I.H., et al. (1994) JNK1: A protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domain. Cell, 76, 1025-1037. doi:10.1016/0092-8674(94)90380-8
[26] Cantley, L.C. (2002) The phosphoinositide 3-kinase pathway. Science, 296, 1655-1657. doi:10.1126/science.296.5573.1655
[27] List of SCADS inhibitor kit III: (2012).
[28] Ushijima, H. and Maeda, M. (2012) cAMP-dependent proteolysis of GATA-6 is linked to JNK-signaling pathway. Biochemical and Biophysical Research Communications, 423, 679-683. doi:10.1016/j.bbrc.2012.06.013
[29] Huggon, I.C., Davies, A., Gove, C., et al. (1997) Molecular cloning of human GATA-6 DNA binding protein: High levels of expression in heart and gut. Biochimica et Biophysica Acta, 1353, 98-102. doi:10.1016/S0167-4781(97)00049-3
[30] Suzuki, E., Evans, T., Lowry, J., et al. (1996) The human GATA-6 gene: Structure, chromosomal location, and regulation of expression by tissue-specific and mitogen-responsive signals. Genomics, 38, 283-290. doi:10.1006/geno.1996.0630
[31] LaVoie, H.A., McCoy, G.L., Blake, C.A. (2004) Expression of the GATA-4 and GATA-6 transcription factors in the fetal rat gonad and in the ovary during postnatal development and pregnancy. Molecular and Cellular Endocrinology, 227, 31-40. doi:10.1016/j.mce.2004.07.016
[32] Visvikis, O., Lorès, P., Boyer, L., et al. (2007) Activated Rac1, but not the tumorigenic variant Rac1b, is ubiquitinated on Lys 147 through a JNK-regulated process. FEBS Journal, 275, 386-396. doi:10.1111/j.1742-4658.2007.06209.x
[33] Chuang, J.Y., Wang, Y.T., Yeh, S.H., et al. (2008) Phos- phorylation by c-Jun NH2-terminal kinase 1 regulates the stability of transcription factor Sp1 during mitosis. Molecular Biology of the Cell, 19, 1139-1151. doi:10.1091/mbc.E07-09-0881
[34] Baker, S.J., Kerppola, T.K., Luk, D., et al. (1992) Jun is phosphorylated by several protein kinases at the same sites that are modified in serum-stimulated fibroblasts. Molecular and Cellular Biology, 12, 4694-4705.
[35] Bousset, K., Oelgeschl?ger, M.H., Henriksson, M., et al. (1994) Regulation of transcription factors c-Myc, Max, and c-Myb by casein kinase II. Cellular & Molecular Biology Research, 40, 501-511.
[36] Sreedhar, A.S., Soti, C. and Csermely, P. (2004) Inhibition of Hsp90: A new strategy for inhibiting protein kinases. Biochimica et Biophysica Acta, 697, 233-242.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.