Proteomic analysis of ACTN4-interacting proteins reveals it’s a putative involvement in mRNA metabolism

Mikhail Khotin, Lidia Turoverova, Vasilisa Aksenova, Nikolai Barlev, Veronika Viktorija Borutinskaite, Alexander Vener, Olga Bajenova, Karl-Eric Magnusson, George P. Pinaev, Dmitri Tentler


Biochemical and Biophysical Research Communications 397 (2010) 192–196


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Alpha-actinin 4 (ACTN4) is an actin-binding protein. In the cytoplasm, ACTN4 participates in structural organisation of the cytoskeleton via cross-linking of actin filaments. Nuclear localisation of ACTN4 has also been reported, but no clear role in the nucleus has been established. In this report, we describe the identification of proteins associated with ACTN4 in the nucleus. A combination of two-dimensional gel electrophoresis (2D-GE) and MALDI-TOF mass-spectrometry revealed a large number of ACTN4-bound proteins that are involved in various aspects of mRNA processing and transport. The association of ACTN4 with different ribonucleoproteins suggests that a major function of nuclear ACTN4 may be regulation of mRNA metabolism and signaling.


[1] K. Honda, T. Yamada, R. Endo, Y. Ino, M. Gotoh, H. Tsuda, Y. Yamada, H. Chiba, S. Hirohashi, Actinin-4, a novel actin-bundling protein associated with cell motility and cancer invasion, J. Cell Biol. 140 (1998) 1383–1393.

[2] A. Bolshakova, O. Petukhova, L. Turoverova, D. Tentler, V. Babakov, K.E. Magnusson, G. Pinaev, Extra-cellular matrix proteins induce re-distribution of alpha-actinin-1 and alpha-actinin-4 in A431 cells, Cell Biol. Int. 3 (2007) 360–365.

[3] V.N. Babakov, O.A. Petukhova, L.V. Turoverova, I.V. Kropacheva, D.G. Tentler, A.V. Bolshakova, E.P. Podolskaya, K.E. Magnusson, G.P. Pinaev, RelA/NFKappaB transcription factor associates with alpha-actinin-4, Exp. Cell Res. 314 (2008) 1030–1038.

[4] A.F. Are, V.E. Galkin, T.V. Pospelova, G.P. Pinaev, The p65/RelA subunit of NFkappaB interacts with actin-containing structures, Exp. Cell Res. 256 (2000) 533–544.

[5] V.N. Babakov, D.E. Bobkov, O.A. Petukhova, L.V. Turoverova, I.V. Kropacheva, E.P. Podol’skaya, G.P. Pinaev, Alpha-actinin-4 and p65/RelA subunit of NFkappaB transcription factor are co-localized and migrate together into the nucleus in EGF-stimulated A431 cell, Tsitologia 46 (2004) 1064–1072.

[6] S. Goffart, A. Franko, C.S. Clemen, R.J. Wiesne, Alpha-actinin 4 and BAT1 interaction with the cytochrome c promoter upon skeletal muscle differentiation, Curr. Genet. 49 (2006) 125–135.

[7] M.T. Poch, L. A1-Kassim, S.M. Smolinski, R.N. Hines, Two distinct classes of CCAAT box elements that bind nuclear factor-Y. Alpha-actinin-4: potential role in human CYP1A1 regulation, Toxicol. Appl. Pharmacol. 199 (2004) 239–250.

[8] S. Chakraborty, E.L. Reineke, M. Lam, X. Li, Y. Liu, C. Gao, S. Khurana, H.Y. Kao, Alpha-actinin 4 potentiates myocyte enhancer factor-2 transcription activity by antagonizing histone deacetylase 7, J. Biol. Chem. 281 (2006) 35070–35080.

[9] U.K. Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature 227 (1970) 680–685.

[10] H. Towbin, T. Staehelin, J. Gordon, Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications, Proc. Natl. Acad. Sci. USA 76 (1979) 4350–4354.

[11] A. Shevchenko, H. Tomas, J. Havlis, J.V. Olsen, M. Mann, In-gel digestion for mass spectrometric characterization of proteins and proteomes, Nat. Prot. 1 (2006) 2856–2860.

[12] S.A. Akker, S. Misra, S. Aslam, E.L. Morgan, P.J. Smith, B. Khoo, S.L. Chew, Prespliceosomal binding of U1 small nuclear ribonucleoprotein (RNP) and heterogenous nuclear RNP E1 is associated with suppression of a growth hormone receptor pseudoexon, Mol. Endocrinol. 21 (2007) 2529–2540.

[13] Q. Meng, S.K. Rayala, A.E. Gururaj, A.H. Talukder, B.W. O’Malley, R. Kumar, Signaling-dependent and coordinated regulation of transcription, splicing, and translation resides in a single coregulator, PCBP1, Proc. Natl. Acad. Sci. USA 107 (2007) 5866–5871.

[14] E. Buratti, F.E. Baralle, Characterization and functional implications of the RNA binding properties of nuclear factor TDP-43, a novel splicing regulator of CFTR exon 9, J. Biol. Chem. 276 (2001) 36337–36343.

[15] C.A. Neumann, D.S. Krause, C.V. Carman, S. Das, D.P. Dubey, J.L. Abraham, R.T. Bronson, Y. Fujiwara, S.H. Orkin, R.A. Van Etten, Essential role for the peroxiredoxin Prdx1 in erythrocyte antioxidant defence and tumour suppression, Nature 424 (2003) 561–565.

[16] E.A. Papakonstanti, C. Stournaras, Cell responses regulated by early reorganization of actin cytoskeleton, FEBS Lett. 582 (2008) 2120–2127.

[17] T. Hara, K. Honda, M. Shitashige, M. Ono, H. Matsuyama, K. Naito, S. Hirohashi, T. Yamada, Mass spectrometry analysis of the native protein complex containing actinin-4 in prostate cancer cells, Mol. Cell. Proteomics 6 (2007) 479–491.

[18] M. Kumeta, S.H. Yoshimura, M. Harata, K. Takeyasu, Molecular mechanisms underlying nucleocytoplasmic shuttling of actinin-4, J. Cell. Sci. 123 (2010) 1020–1030.

[19] T. Pederson, U. Aebi, Actin in the nucleus: what form and what for?, J Struct. Biol. 40 (2002) 3–9.

[20] P. Percipalle, A. Jonsson, D. Nashchekin, C. Karlsson, T. Bergman, A. Guialis, B. Daneholt, Nuclear actin is associated with a specific subset of hnRNP A/B-type proteins, Nucl. Acids Res. 30 (2002) 1725–1734.

[21] X. Zhu, X. Zeng, B. Huang, S. Hao, Actin is closely associated with RNA polymerase II and involved in activation of gene transcription, Biochem. Biophys. Res. Commun. 321 (2004) 623–630.

[22] V.V. Philimonenko, J. Zhao, S. Iben, H. Dingova, K. Kysela, M. Kahle, H. Zentgraf, W.A. Hofmann, P. de Lanerolle, P. Hozak, I. Grummt, Nuclear actin and myosin I are required for RNA polymerase I transcription, Nat. Cell Biol. 6 (2004) 1165–1172.

[23] A. Kukalev, Y. Nord, C. Palmberg, T. Bergman, P. Percipalle, Actin and hnRNP U cooperate for productive transcription by RNA polymerase II, Nat. Struct. Mol. Biol. 12 (2005) 238–244.

[24] W.A. Hofmann, L. Stojiljkovic, B. Fuchsova, G.M. Vargas, E. Mavrommatis, V. Philimonenko, K. Kysela, J.A. Goodrich, J.L. Lessard, T.J. Hope, P. Hozak, P. De Lanerolle, Actin is part of pre-initiation complexes and is necessary for transcription by RNA polymerase II, Nat. Cell Biol. 6 (2004) 1094–1101.

[25] Y. He, R. Smith, Nuclear functions of heterogeneous nuclear ribonucleoproteins A/B, Cell. Mol. Life Sci. 66 (2009) 1239–1256.

[26] A.M. Krecic, M.S. Swanson, HnRNP complexes: composition, structure, and function, Curr. Opin. Cell Biol. 11 (1999) 363–371.

[27] C. Jeronimo, D. Forget, A. Bouchard, Q. Li, G. Chua, C. Poitras, C. Therien, D. Bergeron, S. Bourassa, J. Greenblatt, B. Chabot, G.G. Poirier, T.R. Hughes, M. Blanchette, D.H. Price, B. Coulombe, Systematic analysis of the protein interaction network for the human transcription machinery reveals the identity of the 7SK capping enzyme, Mol. Cell. 27 (2007) 262–274.

[28] J. Nielsen, M.A. Kristensen, M. Willemoës, F.C. Nielsen, J. Christiansen, Sequential dimerization of human zipcode-binding protein IMP1 on RNA: a cooperative mechanism providing RNP stability, Nucl. Acids Res. 32 (2004) 4368–4376.

[29] L. Jonson, J. Vikesaa, A. Krogh, L.K. Nielsen, T. Hansen, R. Borup, A.H. Johnsen, J. Christiansen, F.C. Nielsen, Molecular composition of IMP1 ribonucleoprotein granules, Mol. Cell. Proteomics 6 (2007) 798–811.

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