Abstract
The p53 tumor suppressor protein is widely known for its role as a sequence-specific transcription factor that regulates the expression of stress response genes. Here, we report the identification of LIMK2, which encodes a kinase that regulates actin dynamics through phosphorylation of cofilin, as a p53 target upregulated by DNA damage. Interestingly, the splice variant LIMK2b, but not LIMK2a, was induced in a p53-dependent manner through an intronic consensus p53-binding site. Depletion of LIMK2b leads to early exit of G2/M arrest after DNA damage, whereas its overexpression prolongs the arrest. These responses are recapitulated by ectopic expression of the active cofilin S3A mutant and the inactive cofilin S3D mutant, respectively, suggesting that LIMK2b may modulate G2/M arrest through cofilin phosphorylation. Furthermore, in support of its potential role as a tumor suppressor, LIMK2b was downregulated in esophageal and thyroid cancers, as well as in a number of established cancer cell lines, and its expression suppresses cancer cell migration. Taken together, our results unveil a novel pathway whereby LIMK2b, acting downstream of p53, ensures proper execution of checkpoint arrest by modulating the dynamics of actin polymerization.
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References
Abe H, Obinata T, Minamide LS, Bamburg JR . (1996). Xenopus laevis actin-depolymerizing factor/cofilin: a phosphorylation-regulated protein essential for development. J Cell Biol 132: 871–885.
Amano T, Kaji N, Ohashi K, Mizuno K . (2002). Mitosis-specific activation of LIM motif-containing protein kinase and roles of cofilin phosphorylation and dephosphorylation in mitosis. J Biol Chem 277: 22093–22102.
Amano T, Tanabe K, Eto T, Narumiya S, Mizuno K . (2001). LIM-kinase 2 induces formation of stress fibres, focal adhesions and membrane blebs, dependent on its activation by Rho-associated kinase-catalysed phosphorylation at threonine-505. Biochem J 354: 149–159.
Arber S, Barbayannis FA, Hanser H, Schneider C, Stanyon CA, Bernard O et al. (1998). Regulation of actin dynamics through phosphorylation of cofilin by LIM-kinase. Nature 393: 805–809.
Bode AM, Dong Z . (2004). Post-translational modification of p53 in tumorigenesis. Nat Rev Cancer 4: 793–805.
Croft DR, Olson MF . (2006). The Rho GTPase effector ROCK regulates cyclin A, cyclin D1, and p27Kip1 levels by distinct mechanisms. Mol Cell Biol 26: 4612–4627.
Gadéa G, de Toledo M, Anguille C, Roux P . (2007). Loss of p53 promotes RhoA-ROCK-dependent cell migration and invasion in 3D matrices. J Cell Biol 178: 23–30.
Gadéa G, Lapasset L, Gauthier-Rouvière C, Roux P . (2002). Regulation of Cdc42-mediated morphological effects: a novel function for p53. EMBO J 21: 2373–2382.
Goyal P, Pandey D, Behring A, Siess W . (2005). Inhibition of nuclear import of LIMK2 in endothelial cells by protein kinase C-dependent phosphorylation at Ser-283. J Biol Chem 280: 27569–27577.
Gunsalus KC, Bonaccorsi S, Williams E, Verni F, Gatti M, Goldberg ML . (1995). Mutations in twinstar, a Drosophila gene encoding a cofilin/ADF homologue, result in defects in centrosome migration and cytokinesis. J Cell Biol 131: 1243–1259.
Guo F, Gao Y, Wang L, Zheng Y . (2003). p19ARF-p53 tumor suppressor pathway regulates cell motility by suppression of phosphoinositide 3-kinase and Rac1 GTPase activities. J Biol Chem 278: 14414–14419.
Harms K, Nozell S, Chen X . (2004). The common and distinct target genes of the p53 family transcription factors. Cell Mol Life Sci 61: 822–842.
Hotulainen P, Paunola E, Vartiainen MK, Lappalainen P . (2005). Actin-depolymerizing factor and cofilin-1 play overlapping roles in promoting rapid F-actin depolymerization in mammalian nonmuscle cells. Mol Biol Cell 16: 649–664.
Ikebe C, Ohashi K, Fujimori T, Bernard O, Noda T, Robertson EJ et al. (1997). Mouse LIM-kinase 2 gene: cDNA cloning, genomic organization, and tissue-specific expression of two alternatively initiated transcripts. Genomics 46: 504–508.
Jetten AM . (2009). Retinoid-related orphan receptors (RORs):critical roles in development, immunity, circadian rhythm, and cellular metabolism. Nucl Recept Signal 7: e003.
Lavin MF, Gueven N . (2006). The complexity of p53 stabilization and activation. Cell Death Differ 13: 941–950.
Lee S, Helfman DM . (2004). Cytoplasmic p21Cip1 is involved in Ras-induced inhibition of the ROCK/LIMK/Cofilin pathway. J Biol Chem 279: 1885–1891.
Li Z, Wang C, Jiao X, Lu W, Fu M, Quong AA et al. (2006). Cyclin D1 regulates cellular migration through the inhibition of thrombospondin 1 and ROCK signaling. Mol Cell Biol 26: 4240–4256.
Lu Y, Yi Y, Liu P, Wen W, James M, Wang D et al. (2007). Common human cancer genes discovered by integrated gene-expression analysis. PLoS ONE 2: e1149.
Maekawa M, Ishizaki T, Boku S, Watanabe N, Fujita A, Iwamatsu A et al. (1999). Signaling from Rho to the actin cytoskeleton through protein kinases ROCK and LIM-kinase. Science 285: 895–898.
Murray-Zmijewski F, Slee EA, Lu X . (2008). A complex barcode underlies the heterogeneous response of p53 to stress. Nat Rev Mol Cell Biol 9: 702–712.
Nomoto S, Tatematsu Y, Takahashi T, Osada H . (1999). Cloning and characterization of the alternative promoter regions of the human LIMK2 gene responsible for alternative transcripts with tissue-specific expression. Gene 236: 259–271.
Ohashi K, Nagata K, Maekawa M, Ishizaki T, Narumiya S, Mizuno K . (2000). Rho-associated kinase ROCK activates LIM-kinase 1 by phosphorylation at threonine 508 within the activation loop. J Biol Chem 275: 3577–3582.
Okano I, Hiraoka J, Otera H, Nunoue K, Ohashi K, Iwashita S et al. (1995). Identification and characterization of a novel family of serine/threonine kinases containing two N-terminal LIM motifs. J Biol Chem 270: 31321–31330.
Olivier M, Eeles R, Hollstein M, Khan MA, Harris CC, Hainaut P . (2002). The IARC TP53 database: new online mutation analysis and recommendations to users. Hum Mutat 19: 607–614.
Olsson A, Manzl C, Strasser A, Villunger A . (2007). How important are post-translational modifications in p53 for selectivity in target-gene transcription and tumour suppression? Cell Death Differ 14: 1561–1575.
Ongusaha PP, Kim HG, Boswell SA, Ridley AJ, Der CJ, Dotto GP et al. (2006). RhoE is a pro-survival p53 target gene that inhibits ROCK I-mediated apoptosis in response to genotoxic stress. Curr Biol 16: 2466–2472.
Osada H, Hasada K, Inazawa J, Uchida K, Ueda R, Takahashi T et al. (1996). Subcellular localization and protein interaction of the human LIMK2 gene expressing alternative transcripts with tissue-specific regulation. Biochem Biophys Res Commun 229: 582–589.
Ou Y-H, Chung P-H, Hsu F-F, Sun T-P, Chang W-Y, Shieh S-Y . (2007). The candidate tumor suppressor BTG3 is a transcriptional target of p53 that inhibits E2F1. EMBO J 26: 3968–3980.
Riley T, Sontag E, Chen P, Levine A . (2008). Transcriptional control of human p53-regulated genes. Nat Rev Mol Cell Biol 9: 402–412.
Sahai E, Olson MF, Marshall CJ . (2001). Cross-talk between Ras and Rho signaling pathways in transformation favours proliferation and increased motility. EMBO J 20: 755–766.
Scott RW, Olson MF . (2007). LIM kinases: function, regulation and association with human disease. J Mol Med 85: 555–568.
Smolich B, Vo M, Buckley S, Plowman G, Papkoff J . (1997). Cloning and biochemical characterization of LIMK-2, a protein kinase containing two LIM domains. J Biochem 121: 382–388.
Sumi T, Matsumoto K, Nakamura T . (2001). Specific activation of LIM kinase 2 via phosphorylation of threonine 505 by ROCK, a Rho-dependent protein kinase. J Biol Chem 276: 670–676.
Sumi T, Matsumoto K, Takai Y, Nakamura T . (1999). Cofilin phosphorylation and actin cytoskeletal dynamics regulated by Rho- and Cdc42-activated LIM-kinase 2. J Cell Biol 147: 1519–1532.
Takahashi T, Koshimizu U, Abe H, Ohinata T, Nakamura T . (2001). Functional involvement of Xenopus LIM kinases in progression of oocyte maturation. Dev Biol 229: 554–567.
Toledo F, Wahl GM . (2006). Regulating the p53 pathway: in vitro hypotheses, in vivo veritas. Nat Rev Cancer 6: 909–923.
Vardouli L, Moustakas A, Stournaras C . (2005). LIM-kinase 2 and cofilin phosphorylation mediate actin cytoskeleton reorganization induced by transforming growth factor-beta. J Biol Chem 280: 11448–11457.
Zhu Y, McAvoy S, Kuhn R, Smith DI . (2006). RORA, a large common fragile site gene, is involved in cellular stress response. Oncogene 25: 2901–2908.
Acknowledgements
We thank Hsiu-Ting Lin for her expert technical assistance with the RT–PCR. This work was supported by funding from Academia Sinica to S-Y Shieh.
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Hsu, FF., Lin, TY., Chen, JY. et al. p53-Mediated transactivation of LIMK2b links actin dynamics to cell cycle checkpoint control. Oncogene 29, 2864–2876 (2010). https://doi.org/10.1038/onc.2010.40
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DOI: https://doi.org/10.1038/onc.2010.40
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