PROTEIN KINASE D INTERACTS WITH ADAPTOR PROTEIN RUK/CIN85 AND PHOSPHORYLATES IT
DOI: http://dx.doi.org/10.30970/sbi.0303.050
Abstract
Adaptor protein Ruk/CIN85 is built of multiple domains and motives involved in the intermolecular interactions. It functions as a key component of signalling networks that controls important cellular responses in coordinated fashion. The presence of many consensus motives for a number of protein kinases in Ruk/CIN85 structure suggests that phosphorylation is one of the most probable adaptor protein posttranslational modifications capable of regulating its biological activity depending on cellular context. In this study, we have demonstrated that protein kinases D (PKD) 1 and 2 co-immunoprecipitate with Ruk/CIN85 from lysates of HEK293 cells transiently transfected with pRc/CMV-Rukl vector. By using the panel of Ruk/CIN85 GST-fusion fragments for PKD2-Flag precipitation, it was shown that PKD2 interacts with SH3В and, to a less extent, with SH3С Ruk domains, while the simultaneous presence of both domains enhances this interaction. It was also found that PKD2 most strongly interacts with Pro-rich region of Ruk and possesses considerably lower affinity to coil-coiled domain. The results of in vitro kinase assay using anti-PKD2-Flag immunoprecipitates and GST-fusion forms of Ruk SH3 domains, as substrates showed that the highest level of radioactive 32Р incorporation from [γ-32P]ATP was observed in SH3AB fragment comparing with considerably lower intensity of SH3ABC and SH3B phosphorylation. The obtained data suggest that PKD is a novel binding partner forof Ruk/CIN85 involved in the control of its functional activity through phosphorylation.
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1. Aissouni Y., Zapart G., Iovanna J.L. et al. CIN85 regulates the ability of MEKK4 to activate the p38 MAP kinase pathway. Biochem. Biophys. Res. Commun, 2005; 338(2): 808-814. | |
| |
2. Borinstein S.C., Hyatt M.A., Sykes V.W. et al. SETA is a multifunctional adapter protein with three SH3 domains that binds Grb2, Cbl, and the novel SB1 proteins. Cell. Signal, 2000; 12(11): 769-779. | |
| |
3. Borthwick E.B., Korobko I.V., Courtney L. et al. Multiple domains of Ruk/CIN85/SETA/CD2BP3 are involved in interaction with p85α regulatory subunit of PI 3-kinase. J. Mol. Biol, 2004; 343(4): 1135-1146. | |
| |
4. Bowden E.T., Barth M., Thomas D. et al. An invasion-related complex of cortactin, paxillin and PKCmu associates with invadopodia at sites of extracellular matrix degradation. Oncogene, 1999; 18(31): 4440-9. | |
| |
5. Buchman V.L., Luke C., Borthwick E.B. et al. Organization of the mouse Ruk locus and expression of isoforms in mouse tissues. Gene, 2002; 295(1): 13-17. | |
| |
6. Chen B., Borinstein S.C., Gillis J. et al. The glioma-associated protein SETA interacts with AIP1/Alix and ALG-2 and modulates apoptosis in astrocytes. J. Biol. Chem, 2000; 275(25): 19275-19281. | |
| |
7. Dikic I. CIN85/CMS family of adaptor molecules. FEBS Lett, 2002; 529(1): 110-115. | |
| |
8. Dikic I., Giordano S. Negative receptor signaling. Curr. Opin. Cell Biol, 2003; 15(2): 128-35. | |
| |
9. Dustin M.L., Olszowy M.W., Holdorf A.D. et al. A novel adaptor protein orchestrates receptor patterning and cytoskeletal polarity in T-cell contacts. Cell, 1998; 94(5): 667-77. | |
| |
10. Gaidos G., Soni S., Oswald D.J. et al. Structure and function analysis of the CMS/CIN85 protein family identifies actin-bundling properties and heterotypic-complex formation. J. Cell Sci, 2007; 120(Pt 14): 2366-77. | |
| |
11. Gout I., Middleton G., Adu J. et al. Negative regulation of PI 3-kinase by Ruk, a novel adaptor protein. EMBO J., 2000; 19(15): 4015-4025. | |
| |
12. Haglund K., Shimokawa N., Szymkiewicz I., Dikic I. Cbl-directed monoubiquitination of CIN85 is involved in regulation of ligand-induced degradation of EGF receptors. Proc. Natl. Acad. Sci. USA, 2002; 99(19): 12191-12196. | |
| |
13. Havrylov S., Ichioka F., Powell K. et al. Adaptor protein Ruk/CIN85 is associated with a subset of COPI-coated membranes of the Golgi complex. Traffic, 2008; 9(5): 798-812. | |
| |
14. Havrylov S., Rzhepetskyy Y., Malinowska A. et al. Proteins recruited by SH3 domains of Ruk/CIN85 adaptor identified by LC-MS/MS. Proteome Sci, 2009; 7:21-30. | |
| |
15. Ilnytska O.M., Drel' V.R., Shuvayeva H.Yu. et al. Intra- and intermolecular interactions mediated by adaptor protein Ruk/CIN85/SETA..Biopolymers and Cell, 2005; 21(1): 48-54. | |
| |
16. Kit Yu.Ya., Drel V.R., Petriv O.I. et al. Adaptor protein Rukl forms protein-protein complexes with endonuclease activity in HEK293 cells. Biochemistry (Moscow), 2003; 68(7): 810-815. | |
| |
17. Kowanetz K., Husnjak K., Holler D. et al. CIN85 associates with multiple effectors controlling intracellular trafficking of EGF receptors. Mol. Biol. Cell, 2004; 15(7): 3155-3166. | |
| |
18. Kowanetz K., Szymkiewicz I., Haglund K. et al. Identification of a novel proline-arginine motif involved in CIN85-dependent clustering of Cbl and down-regulation of epidermal growth factor receptors. J. Biol. Chem, 2003; 278(41): 39735-39746. | |
| |
19. Kowanetz K., Terzic J. Dikic I. Dab2 links CIN85 with clathrin-mediated receptor internalization. FEBS Lett., 2003; 554(1): 81-87. | |
| |
20. Kurakin A.V., Wu S., Bredesen D.E. Atypical recognition consensus of CIN85/SETA/Ruk SH3 domains revealed by target-assisted iterative screening. J. Biol. Chem, 2003; 278(36): 34102-34109. | |
| |
21. Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 1970; 227(5259): 680-684. | |
| |
22. Liang Y., Kurakin A., Roizman B. Herpes simplex virus 1 infected cell protein 0 forms a complex with CIN85 and Cbl and mediates the degradation of EGF receptor from cell surfaces. Proc. Nat. Acad. Sci. USA, 2005; 102(16): 5838-5843. | |
| |
23. Lynch D.K., Winata S.C., Lyons R.J. et al. A Cortactin-CD2-associated protein (CD2AP) complex provides a novel link between epidermal growth factor receptor endocytosis and the actin cytoskeleton. J Biol Chem, 2003 278(24): 21805-13. | |
| |
24. Mayevska O., Shuvayeva H., Igumentseva N. et al. Expression of adaptor protein Ruk/CIN85 isoforms in cell lines of various tissue origins and human melanoma. Exp. Oncol, 2006; 28(4): 275-81. | |
| |
25. Moarefi I., LaFevre-Bernt M., Sicheri F. et al. Activation of the Src-family tyrosine kinase Hck by SH3 domain displacement. Nature, 1997; 385(6617): 650-3. | |
| |
26. Nam J.M., Onodera Y., Mazaki Y.et al. CIN85, a Cbl-interacting protein, is a component of AMAP1-mediated breast cancer invasion machinery. EMBO J, 2007; 26(3): 647-56. | |
| |
27. Narita T., Nishimura T., Yoshizaki K., Taniyama T. CIN85 associates with TNF receptor 1 via Src and modulates TNF-α-induced apoptosis. Exp. Cell Res, 2005; 304(1): P.256-264. | |
| |
28. Pawson T., Nash P. Assembly of cell regulatory systems through protein interaction domains. Science, 2003; 300(5618): 445-452. | |
| |
29. Peterson G.L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal. Biochem, 1977; 83(2): 346-356. | |
| |
30. Petrelli A., Gilestro G.F., Lanzardo S. et al. The endophilin-CIN85-Cbl complex mediates ligand-dependent downregulation of c-Met. Nature, 2002; 416(6877): 187-190. | |
| |
31. Rozengurt E., Rey O., Waldron R.T. Protein kinase D signaling. J. Biol. Chem, 2005; 280(14): 13205-8. | |
| |
32. Rykx A., De Kimpe L., Mikhalap S. et al. Protein kinase D: a family affair. FEBS Lett, 2003; 546(1): 81-86. | |
| |
33. Sicheri F., Moarefi I., Kuriyan J. Crystal structure of the Src family tyrosine kinase Hck. Nature, 1997; 385(6617): 602-609. | |
| |
34. Soubeyran P., Kowanetz K., Szymkiewicz I. et al. Cbl-CIN85-endophilin complex mediates ligand-induced downregulation of EGF receptors. Nature, 2002; 416(6877): 183-187. | |
| |
35. Tibaldi E.V., Reinherz E.L. CD2BP3, CIN85 and the structurally related adaptor protein CMS bind to the same CD2 cytoplasmic segment, but elicit divergent functional activities. Int. Immunol, 2003; 15(3): 313-329. | |
| |
36. Tossidou I., Kardinal C., Peters I. et al. CD2AP/CIN85 balance determines receptor tyrosine kinase signaling response in podocytes. J. Biol. Chem., 2007; 282(10): 7457-64. | |
| |
37. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc. Natl. Acad. Sci. USA, 1979; 76(9): 4350-4354. | |
| |
38. Verdier F., Valovka T., Zhyvoloup A. et al. Ruk is ubiquitinated but not degraded by the proteasome. Eur. J. Biochem, 2002; 269(14): 3402-3408. | |
| |
39. Watanabe S., Take H., Takeda K. et al. Characterization of the CIN85 adaptor protein and identification of components involved in CIN85 complexes. Biochem. Biophys. Res. Commun, 2000; 278(1): 167-174. | |
| |
40. Williams J.C., Weijland A., Gonfloni S. et al. The 2.35 A crystal structure of the inactivated form of chicken Src: a dynamic molecule with multiple regulatory interactions. J. Mol. Biol, 1997; 274(5): 757-75. | |
| |
41. Xu W., Doshi A., Lei M. et al. Crystal structures of c-Src reveal features of its autoinhibitory mechanism. Mol. Cell, 1999; 3(5): 629-38. | |
| |
42. Yuzava S., Suzuki N.N., Fujioka Y. et al. A molecular mechanism for autoinhibition of the tandem SH3 domains of p47phox, the regulatory subunit of the phagocyte NADPH oxidase. Genes to Cells, 2004; 9(5): 443-456. | |
| |
43. Zhang J., Zheng X., Yang X., Liao K. CIN85 associates with endosomal membrane and binds phosphatidic acid. Cell Res, 2009; 19(6): 733-746. |
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