{"_id":"AAAkeSvhtApjrD7T2","bibbaseid":"xia-levy-levillayer-jia-li-neuveut-buendia-lan-etal-thefourandahalflimonlyprotein2fhl2activatestransforminggrowthfactorbetatgfbetasignalingbyregulatingubiquitinationofthee3ligasearkadia-2013","author_short":["Xia, T.","Levy, L.","Levillayer, F.","Jia, B.","Li, G.","Neuveut, C.","Buendia, M. A.","Lan, K.","Wei, Y."],"bibdata":{"bibtype":"article","type":"article","title":"The four and a half LIM-only protein 2 (FHL2) activates transforming growth factor beta (TGF-beta) signaling by regulating ubiquitination of the E3 ligase Arkadia","volume":"288","issn":"1083-351X (ELECTRONIC); 0021-9258 (LINKING)","shorttitle":"The four and a half LIM-only protein 2 (FHL2) activates transforming growth factor beta (TGF-beta) signaling by regulating ubiquitination of the E3 ligase Arkadia","abstract":"Arkadia is a RING-based ubiquitin ligase that positively regulates TGF-beta signaling by targeting several pathway components for ubiquitination and degradation. However, little is known about the mechanisms controlling Arkadia activity. Here we show that the LIM-only protein FHL2 binds and synergistically cooperates with Arkadia to activate Smad3/Smad4-dependent transcription. Knockdown of FHL2 by RNA interference decreases Arkadia level and restricts the amplitude of Arkadia-induced TGF-beta target gene responses. We found that Arkadia is ubiquitinated via K63- and K27-linked polyubiquitination. A single mutation at the RING domain that abolishes the E3 activity diminishes Arkadia ubiquitination, indicating that this modification partly involves autocatalytic process. Mutation of seven lysines at the C-terminal region of Arkadia severely impairs ubiquitination through the K27 but not the K63 linkage and slows down the turnover of Arkadia, suggesting that K27-linked polyubiquitination might promote proteolysis-dependent regulation of Arkadia. We show that FHL2 increases the half-life of Arkadia through inhibition of ubiquitin chain assembly on the protein, which provides a molecular basis for functional cooperation between Arkadia and FHL2 in enhancing TGF-beta signaling. Our study uncovers a novel regulatory mechanism of Arkadia by ubiquitination and identifies FHL2 as important regulator of Arkadia ubiquitination and TGF-beta signal transduction.","number":"3","journal":"J Biol Chem","author":[{"propositions":[],"lastnames":["Xia"],"firstnames":["T."],"suffixes":[]},{"propositions":[],"lastnames":["Levy"],"firstnames":["L."],"suffixes":[]},{"propositions":[],"lastnames":["Levillayer"],"firstnames":["F."],"suffixes":[]},{"propositions":[],"lastnames":["Jia"],"firstnames":["B."],"suffixes":[]},{"propositions":[],"lastnames":["Li"],"firstnames":["G."],"suffixes":[]},{"propositions":[],"lastnames":["Neuveut"],"firstnames":["C."],"suffixes":[]},{"propositions":[],"lastnames":["Buendia"],"firstnames":["M.","A."],"suffixes":[]},{"propositions":[],"lastnames":["Lan"],"firstnames":["K."],"suffixes":[]},{"propositions":[],"lastnames":["Wei"],"firstnames":["Y."],"suffixes":[]}],"month":"January","year":"2013","keywords":"Animals Binding Sites Cell Line, Reporter Half-Life Humans LIM-Homeodomain Proteins/ genetics/metabolism Luciferases Mice Muscle Proteins/ genetics/metabolism Mutation Nuclear Proteins/ genetics/metabolism Protein Binding Protein Structure, Tertiary Signal Transduction Transcription Factors/ genetics/metabolism Transfection Transforming Growth Factor beta/ genetics/metabolism Ubiquitin/ genetics/metabolism Ubiquitin-Protein Ligases/ genetics/metabolism Ubiquitination, Tumor Fibroblasts/cytology/metabolism Gene Expression Regulation Genes","pages":"1785–94","bibtex":"@article{xia_four_2013,\n\ttitle = {The four and a half {LIM}-only protein 2 ({FHL2}) activates transforming growth factor beta ({TGF}-beta) signaling by regulating ubiquitination of the {E3} ligase {Arkadia}},\n\tvolume = {288},\n\tissn = {1083-351X (ELECTRONIC); 0021-9258 (LINKING)},\n\tshorttitle = {The four and a half {LIM}-only protein 2 ({FHL2}) activates transforming growth factor beta ({TGF}-beta) signaling by regulating ubiquitination of the {E3} ligase {Arkadia}},\n\tabstract = {Arkadia is a RING-based ubiquitin ligase that positively regulates TGF-beta signaling by targeting several pathway components for ubiquitination and degradation. However, little is known about the mechanisms controlling Arkadia activity. Here we show that the LIM-only protein FHL2 binds and synergistically cooperates with Arkadia to activate Smad3/Smad4-dependent transcription. Knockdown of FHL2 by RNA interference decreases Arkadia level and restricts the amplitude of Arkadia-induced TGF-beta target gene responses. We found that Arkadia is ubiquitinated via K63- and K27-linked polyubiquitination. A single mutation at the RING domain that abolishes the E3 activity diminishes Arkadia ubiquitination, indicating that this modification partly involves autocatalytic process. Mutation of seven lysines at the C-terminal region of Arkadia severely impairs ubiquitination through the K27 but not the K63 linkage and slows down the turnover of Arkadia, suggesting that K27-linked polyubiquitination might promote proteolysis-dependent regulation of Arkadia. We show that FHL2 increases the half-life of Arkadia through inhibition of ubiquitin chain assembly on the protein, which provides a molecular basis for functional cooperation between Arkadia and FHL2 in enhancing TGF-beta signaling. Our study uncovers a novel regulatory mechanism of Arkadia by ubiquitination and identifies FHL2 as important regulator of Arkadia ubiquitination and TGF-beta signal transduction.},\n\tnumber = {3},\n\tjournal = {J Biol Chem},\n\tauthor = {Xia, T. and Levy, L. and Levillayer, F. and Jia, B. and Li, G. and Neuveut, C. and Buendia, M. A. and Lan, K. and Wei, Y.},\n\tmonth = jan,\n\tyear = {2013},\n\tkeywords = {Animals Binding Sites Cell Line, Reporter Half-Life Humans LIM-Homeodomain Proteins/ genetics/metabolism Luciferases Mice Muscle Proteins/ genetics/metabolism Mutation Nuclear Proteins/ genetics/metabolism Protein Binding Protein Structure, Tertiary Signal Transduction Transcription Factors/ genetics/metabolism Transfection Transforming Growth Factor beta/ genetics/metabolism Ubiquitin/ genetics/metabolism Ubiquitin-Protein Ligases/ genetics/metabolism Ubiquitination, Tumor Fibroblasts/cytology/metabolism Gene Expression Regulation Genes},\n\tpages = {1785--94},\n}\n\n\n\n","author_short":["Xia, T.","Levy, L.","Levillayer, F.","Jia, B.","Li, G.","Neuveut, C.","Buendia, M. A.","Lan, K.","Wei, Y."],"key":"xia_four_2013","id":"xia_four_2013","bibbaseid":"xia-levy-levillayer-jia-li-neuveut-buendia-lan-etal-thefourandahalflimonlyprotein2fhl2activatestransforminggrowthfactorbetatgfbetasignalingbyregulatingubiquitinationofthee3ligasearkadia-2013","role":"author","urls":{},"keyword":["Animals Binding Sites Cell Line","Reporter Half-Life Humans LIM-Homeodomain Proteins/ genetics/metabolism Luciferases Mice Muscle Proteins/ genetics/metabolism Mutation Nuclear Proteins/ genetics/metabolism Protein Binding Protein Structure","Tertiary Signal Transduction Transcription Factors/ genetics/metabolism Transfection Transforming Growth Factor beta/ genetics/metabolism Ubiquitin/ genetics/metabolism Ubiquitin-Protein Ligases/ genetics/metabolism Ubiquitination","Tumor Fibroblasts/cytology/metabolism Gene Expression Regulation Genes"],"metadata":{"authorlinks":{}}},"bibtype":"article","biburl":"http://bibbase.org/zotero/biblioial","dataSources":["ftoP3zPyb2N3b9Noc"],"keywords":["animals binding sites cell line","reporter half-life humans lim-homeodomain proteins/ genetics/metabolism luciferases mice muscle proteins/ genetics/metabolism mutation nuclear proteins/ genetics/metabolism protein binding protein structure","tertiary signal transduction transcription factors/ genetics/metabolism transfection transforming growth factor beta/ genetics/metabolism ubiquitin/ genetics/metabolism ubiquitin-protein ligases/ genetics/metabolism ubiquitination","tumor fibroblasts/cytology/metabolism gene expression regulation genes"],"search_terms":["four","half","lim","protein","fhl2","activates","transforming","growth","factor","beta","tgf","beta","signaling","regulating","ubiquitination","ligase","arkadia","xia","levy","levillayer","jia","li","neuveut","buendia","lan","wei"],"title":"The four and a half LIM-only protein 2 (FHL2) activates transforming growth factor beta (TGF-beta) signaling by regulating ubiquitination of the E3 ligase Arkadia","year":2013}