Ukr.Biochem.J. 2014; Volume 86, Issue 5, Sep-Oct, pp. 102-110

doi: https://doi.org/10.15407/ubj86.05.102

Multiple molecular forms of adaptor protein Ruk/CIN85 specifically associate with different subcellular compartments in human breast adenocarcinoma MCF-7 cells

26.06.2015   Uncategorized   No comments

B. O. Vynnytska-Myronovska1, Ya. P. Bobak1, G. V. Pasichnyk2,
N. I. Igumentseva1, A. A. Samoylenko2, L. B. Drobot2

1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: drobot@biochem.kiev.ua

Ruk/CIN85 is a receptor-proximal ‘signalling’ adaptor that possesses three SH3 domains, Pro- and Ser-rich regions and C-terminal coiled-coil domain. It employs distinct domains and motifs to act as a transducer platform in intracellular signalling. Based on cDNA analysis, various isoforms of Ruk/CIN85 with different combination of protein-protein interaction domains as well as additional Ruk/CIN85 forms that are the products of post-translational modifications have been demonstrated. Nevertheless, there is no precise information regarding both the subcellular distribution and the role of Ruk/CIN85 multiple molecular forms in cellular responses. Using MCF-7 human breast adenocarcinoma cells and cell fractionation technique, specific association of Ruk/CIN85 molecular forms with different subcellular compartments was demonstrated. Induction of apoptosis of MCF-7 cells by doxorubicin treatment or by serum deprivation resulted in the system changes of Ruk/CIN85 molecular forms intracellular localization as well as their ratio. The data obtained provide a new insight into potential physiological significance of Ruk/CIN85 molecular forms in the regulation of various cellular functions.

Keywords: , , , , ,

References:

  1. Levchenko A, Bruck J, Sternberg PW. Scaffold proteins may biphasically affect the levels of mitogen-activated protein kinase signaling and reduce its threshold properties. Proc Natl Acad Sci U S A. 2000 May 23;97(11):5818-23. PubMed, PubMedCentral, CrossRef
  2. Pawson T. Dynamic control of signaling by modular adaptor proteins. Curr Opin Cell Biol. 2007 Apr;19(2):112-6. Epub 2007 Feb 20. Review. PubMed, CrossRef
  3. Gout I, Middleton G, Adu J, Ninkina NN, Drobot LB, Filonenko V, Matsuka G, Davies AM, Waterfield M, Buchman VL. Negative regulation of PI 3-kinase by Ruk, a novel adaptor protein. EMBO J. 2000 Aug 1;19(15):4015-25. PubMed,  PubMedCentral, CrossRef
  4. Take H, Watanabe S, Takeda K, Yu ZX, Iwata N, Kajigaya S. Cloning and characterization of a novel adaptor protein, CIN85, that interacts with c-Cbl. Biochem Biophys Res Commun. 2000 Feb 16;268(2):321-8. PubMed, CrossRef
  5. Hutchings NJ, Clarkson N, Chalkley R, Barclay AN, Brown MH. Linking the T cell surface protein CD2 to the actin-capping protein CAPZ via CMS and CIN85. J Biol Chem. 2003 Jun 20;278(25):22396-403. Epub 2003 Apr 10. PubMed, CrossRef
  6. Schmidt MH, Chen B, Randazzo LM, Bogler O. SETA/CIN85/Ruk and its binding partner AIP1 associate with diverse cytoskeletal elements, including FAKs, and modulate cell adhesion. J Cell Sci. 2003 Jul 15;116(Pt 14):2845-55. Epub 2003 May 27. PubMed, CrossRef
  7. Tibaldi EV, Reinherz EL. CD2BP3, CIN85 and the structurally related adaptor protein CMS bind to the same CD2 cytoplasmic segment, but elicit divergent functional activities. Int Immunol. 2003 Mar;15(3):313-29. PubMed, CrossRef
  8. Nam JM, Onodera Y, Mazaki Y, Miyoshi H, Hashimoto S, Sabe H. CIN85, a Cbl-interacting protein, is a component of AMAP1-mediated breast cancer invasion machinery. EMBO J. 2007 Feb 7;26(3):647-56. Epub 2007 Jan 25. PubMed, PubMedCentral, CrossRef
  9. Havrylov S, Redowicz MJ, Buchman VL. Emerging roles of Ruk/CIN85 in vesicle-mediated transport, adhesion, migration and malignancy. Traffic. 2010 Jun;11(6):721-31. Epub 2010 Mar 17. Review. PubMed, CrossRef
  10. Samoylenko A., Vynnytska-Myronovska B., Byts N., Kozlova N., Basaraba O., Pasichnyk G., Palyvoda K., Bobak Y., Barska M., Mayevska O., Rzhepetsky Y., Shuvayeva H., Lyzogubov V., Usenko V., Savran V., Volodko N., Buchman V., Kietzmann T, Drobot L. Increased levels of the HER1 adaptor protein Rukl/CIN85 contribute to breast cancer malignancy. Carcinogenesis. 2012 Oct;33(10):1976-1984. PubMed, CrossRef
  11. Chen B, Borinstein SC, Gillis J, Sykes VW, Bogler O. The glioma-associated protein SETA interacts with AIP1/Alix and ALG-2 and modulates apoptosis in astrocytes. J Biol Chem. 2000 Jun 23;275(25):19275-81. PubMed, CrossRef
  12. Narita T, Nishimura T, Yoshizaki K, Taniyama T. CIN85 associates with TNF receptor 1 via Src and modulates TNF-alpha-induced apoptosis. Exp Cell Res. 2005 Mar 10;304(1):256-64. Epub 2004 Dec 1. PubMed, CrossRef
  13. Petrelli A, Gilestro GF, Lanzardo S, Comoglio PM, Migone N, Giordano S. The endophilin-CIN85-Cbl complex mediates ligand-dependent downregulation of c-Met. Nature. 2002 Mar 14;416(6877):187-90. PubMed, CrossRef
  14. Soubeyran P, Kowanetz K, Szymkiewicz I, Langdon WY, Dikic I. Cbl-CIN85-endophilin complex mediates ligand-induced downregulation of EGF receptors. Nature. 2002 Mar 14;416(6877):183-7. Erratum in: Nature 2002 May 2;417(6884):102. PubMed, CrossRef
  15. Havrylov S, Ichioka F, Powell K, Borthwick EB, Baranska J, Maki M, Buchman VL. Adaptor protein Ruk/CIN85 is associated with a subset of COPI-coated membranes of the Golgi complex. Traffic. 2008 May;9(5):798-812. Epub 2008 Feb 11. PubMed, CrossRef
  16. Narita T, Ando A, Mikami Y, Taniyama T. Overexpression of CIN85 suppresses the growth of herpes simplex virus in HeLa cells. Exp Cell Res. 2005 Dec 10;311(2):265-71. Epub 2005 Oct 11. PubMed, CrossRef
  17. Buchman VL, Luke C, Borthwick EB, Gout I, Ninkina N. Organization of the mouse Ruk locus and expression of isoforms in mouse tissues. Gene. 2002 Jul 24;295(1):13-17. PubMed, CrossRef
  18. Narita T, Amano F, Yoshizaki K, Nishimoto N, Nishimura T, Tajima T, Namiki H, Taniyama T. Assignment of SH3KBP1 to human chromosome band Xp22.1–>p21.3 by in situ hybridization. Cytogenet Cell Genet. 2001;93(1-2):133-4. PubMed
  19. Finniss S, Movsisyan A, Billecke C, Schmidt M, Randazzo L, Chen B, Bögler O. Studying protein isoforms of the adaptor SETA/CIN85/Ruk with monoclonal antibodies. Biochem Biophys Res Commun. 2004 Dec 3;325(1):174-82. PubMed, CrossRef
  20. Borthwick E. B., Korobko I. V., Luke C., Drel V. R., Fedyshyn Y. Y., Ninkina N., Drobot L. B., Buchman V. L. 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. PubMed, CrossRef
  21. 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 U S A. 2002 Sep 17;99(19):12191-6. Epub 2002 Sep 6. PubMed,  PubMedCentral, CrossRef
  22. Verdier F, Valovka T, Zhyvoloup A, Drobot LB, Buchman V, Waterfield M, Gout I. Ruk is ubiquitinated but not degraded by the proteasome. Eur J Biochem. 2002 Jul;269(14):3402-8. PubMed, CrossRef
  23. Tossidou I, Niedenthal R, Klaus M, Teng B, Worthmann K, King BL, Peterson KJ, Haller H, Schiffer M. CD2AP regulates SUMOylation of CIN85 in podocytes. Mol Cell Biol. 2012 Mar;32(6):1068-79. Epub 2011 Dec 27. PubMed, PubMedCentral, CrossRef
  24. Rzepetskyi Y. А., Samoylenko A. A., Kucha­renko O. P., Mikhalap S. V., Sidorenko S. P., Hausser A., Drobot L. B. Protein kinase D interacts with adaptor protein Ruk/CIN85 and phosphorylates it. Studia Biologica. 2009;3(3):17‑28.
  25. Bior BK, Ballif BA. Dab1 stabilizes its interaction with Cin85 by suppressing Cin85 phosphorylation at serine 587. FEBS Lett. 2013 Jan 4;587(1):60-6. Epub 2012 Nov 21. PubMed, PubMedCentral, CrossRef
  26. Schroeder B, Srivatsan S, Shaw A, Billadeau D, McNiven MA. CIN85 phosphorylation is essential for EGFR ubiquitination and sorting into multivesicular bodies. Mol Biol Cell. 2012 Sep;23(18):3602-11. Epub 2012 Jul 25. PubMed,  PubMedCentral,  CrossRef
  27. Hallberg E., Kihlmark M. Nucleus and nuclear envelope: methods for preparation. ELS: Nature Publ Group, 2001.
  28. Mayevska O, Shuvayeva H, Igumentseva N, Havrylov S, Basaraba O, Bobak Y, Barska M, Volod’ko N, Baranska J, Buchman V, Drobot L. Expression of adaptor protein Ruk/CIN85 isoforms in cell lines of various tissue origins and human melanoma. Exp Oncol. 2006 Dec;28(4):275-81. PubMed
  29. Swarup G., Radha V. Nuclear matrix: methods of preparation. ELS: Nature Publ. Group, 2001.
  30. Kagawa S, Gu J, Honda T, McDonnell TJ, Swisher SG, Roth JA, Fang B. Deficiency of caspase-3 in MCF7 cells blocks Bax-mediated nuclear fragmentation but not cell death. Clin Cancer Res. 2001 May;7(5):1474-80. PubMed

 

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