Abstract
Rock proteins are Rho GTPase-dependent serine/threonine kinases with crucial roles in F-actin dynamics and cell transformation. By analogy with other protein kinase families, it can be assumed that Rock proteins act, at least in part, through the regulation of gene expression events. However, with the exception of some singular transcriptional targets recently identified, the actual impact of these kinases on the overall cell transcriptome remains unknown. To address this issue, we have used a microarray approach to compare the transcriptomes of exponentially growing NIH3T3 cells that had been untreated or treated with Y27632, a well known specific inhibitor for Rock kinase activity. We show here that the Rock pathway promotes a weak impact on the fibroblast transcriptome, since its inhibition only results in changes in the expression of 2.3% of all the genes surveyed in the microarrays. Most Y27632-dependent genes are downregulated at moderate levels, indicating that the Rock pathway predominantly induces the upregulation of transcriptionally active genes. Although functionally diverse, a common functional leitmotiv of Y27632-dependent genes is the implication of their protein products in cytoskeletal-dependent processes. Taken together, these results indicate that Rock proteins can modify cytoskeletal dynamics by acting at post-transcriptional and transcriptional levels. In addition, they suggest that the main target of these serine/threonine kinases is the phosphoproteome and not the transcriptome.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Bustelo XR, Sauzeau V, Berenjeno IM (2007) GTP-binding proteins of the Rho/Rac family: regulation, effectors and functions in vivo. Bioessays. Apr 29:356–370
Van Aelst L, D’souza-Schorey C (1997) Rho GTPases and signaling networks. Genes Dev 11:2295–2322.
Symons M, Rusk N (2003) Control of vesicular trafficking by Rho GTPases. Curr Biol;13:R409–418.
Etienne-Manneville S, Hall A (2002) Rho GTPases in cell biology. Nature 420:629–635.
Coleman ML, Marshall CJ, Olson MF (2004) RAS and RHO GTPases in G1-phase cell-cycle regulation. Nat Rev Mol Cell Biol 5:355–366.
Jaffe AB, Hall A (2005) Rho GTPases: biochemistry and biology. Annu Rev Cell Dev Biol 21:247–269.
Colicelli J (2004) Human RAS superfamily proteins and related GTPases (250)RE13.
DerMardirossian C, Bokoch GM (2005) GDIs: central regulatory molecules in Rho GTPase activation. Trends Cell Biol 15:356–363.
Dransart E, Olofsson B, Cherfils J (2005) RhoGDIs revisited: novel roles in Rho regulation. Traffic 6:957–966.
Rossman KL, Der CJ, Sondek J (2005) GEF means go: turning on RHO GTPases with guanine nucleotide-exchange factors. Nat Rev Mol Cell Biol 6:167–180.
Bos JL, Rehmann H, Wittinghofer A (2007) GEFs and GAPs: critical elements in the control of small G proteins. Cell 129:865–877.
Benitah SA, Valeron PF, van Aelst L, Marshall CJ, Lacal JC (2004) Rho GTPases in human cancer: an unresolved link to upstream and downstream transcriptional regulation. Biochim Biophys Acta 1705:121–132.
Boettner B, Van Aelst L (2002) The role of Rho GTPases in disease development. Gene 286:155–174.
Budzyn K, Marley PD, Sobey CG (2002) Targeting Rho and Rho-kinase in the treatment of cardiovascular disease. Trends Pharmacol Sci 27:97–104.
Sahai E, Marshall CJ (2002) RHO-GTPases and cancer. Nat Rev Cancer 2:133–142.
Riento K, Ridley AJ (2003) Rocks: multifunctional kinases in cell behaviour. Nat. Rev. Cell Biol 4:446–456.
Mueller BK, Mack H, Teusch N (2005) Rho kinase, a promising drug target for neurological disorders. Nat Rev Drug Discov 4:387–398.
Fu X, Gong MC, Jia T, Somlyo AV, Somlyo AP (1998) The effects of the Rhokinase inhibitor Y-27632 on arachidonic acid-, GTPgammaS-, and phorbol esterinduced Ca2+-sensitization of smooth muscle. FEBS Lett 440:183–187.
Shirao S, Kashiwagi S, Sato M et al (2002) Sphingosylphosphorylcholine is a novel messenger for Rho-kinase-mediated Ca2+ sensitization in the bovine cerebral artery: unimportant role for protein kinase C. Circulation research 91:112–119.
Riento K, Guasch RM, Garg R, Jin B, Ridley AJ (2003) RhoE binds to ROCK I and inhibits downstream signaling. Mol Cell Biol 23:4219–4229.
Ward Y, Yap SF, Ravichandran V, Matsumura F, Ito M, Spinelli B, Kelly K (2002) The GTP binding proteins Gem and Rad are negative regulators of the Rho-Rho kinase pathway. J Cell Biol 157: 291–302.
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.
Sahai E, Ishizaki T, Narumiya S, Treisman R (1999) Transformation mediated by RhoA requires activity of ROCK kinases. Curr Biol 9:136–145.
Berenjeno IM, Nunez F, Bustelo XR (2007) Transcriptomal profiling of the cellular transformation induced by Rho subfamily GTPases. Oncogene 26:4295–4305.
Chiariello M, Marinissen MJ, Gutkind JS (2001) Regulation of c-myc expression by PDGF through Rho GTPases. Nat Cell Biol 3:580–586.
Han JS, Macarak E, Rosenbloom J, Chung KC, Chaqour B (2003) Regulation of Cyr61/CCN1 gene expression through RhoA GTPase and p38MAPK signaling pathways. Eur J Biochem 270:3408–3421.
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.
Uehata M, Ishizaki T, Satoh H et al (1997) Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature 389:990–994.
Calvano SE, Xiao W, Richards DR et al (2005) A network-based analysis of systemic inflammation in humans. Nature. Oct 13 2005;437(7061):1032–1037.
Coller HA, Sang L, Roberts JM (2006) A new description of cellular quiescence. PLoS Biol 4:e83.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Berenjeno, I.M., Bustelo, X.R. Identification of the Rock-dependent transcriptome in rodent fibroblasts. Clin Transl Oncol 10, 726–738 (2008). https://doi.org/10.1007/s12094-008-0279-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12094-008-0279-5