Myocardin-related transcription factor regulates Nox4 protein expression: linking cytoskeletal orginization to redox state
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Myocardin-related transcription factor regulates Nox4 protein expression : linking cytoskeletal orginization to redox state. / Rozycki, Matthew; Bialik, Janne Folke; Speight, Pam; Dan, Qinghong; Knudsen, Teresa Emmilie Toudal; Szeto, Stephen G.; Yuen, Darren A.; Szászi, Katalin; Pedersen, Stine Helene Falsig; Kapus, András.
I: Journal of Biological Chemistry, Bind 291, Nr. 1, 2016, s. 227-243.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Myocardin-related transcription factor regulates Nox4 protein expression
T2 - linking cytoskeletal orginization to redox state
AU - Rozycki, Matthew
AU - Bialik, Janne Folke
AU - Speight, Pam
AU - Dan, Qinghong
AU - Knudsen, Teresa Emmilie Toudal
AU - Szeto, Stephen G.
AU - Yuen, Darren A.
AU - Szászi, Katalin
AU - Pedersen, Stine Helene Falsig
AU - Kapus, András
N1 - © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2016
Y1 - 2016
N2 - TGFβ-induced expression of the NADPH oxidase Nox4 is essential for fibroblast-myofibroblast transition. Rho has been implicated in Nox4 regulation, but the underlying mechanisms are largely unknown. Myocardin-related transcription factor (MRTF), a Rho/actin polymerization-controlled coactivator of serum response factor, drives myofibroblast transition from various precursors. We have shown that TGFβ is necessary but insufficient for epithelial-myofibroblast transition in intact epithelia; the other prerequisite is the uncoupling of intercellular contacts, which induces Rho-dependent nuclear translocation of MRTF. Because the Nox4 promoter harbors a serum response factor/MRTF cis-element (CC(A/T)6GG box), we asked if MRTF (and thus cytoskeleton organization) could regulate Nox4 expression. We show that Nox4 protein is robustly induced in kidney tubular cells exclusively by combined application of contact uncoupling and TGFβ. Nox4 knockdown abrogates epithelial-myofibroblast transition-associated reactive oxygen species production. Laser capture microdissection reveals increased Nox4 expression in the tubular epithelium also during obstructive nephropathy. MRTF down-regulation/inhibition suppresses TGFβ/contact disruption-provoked Nox4 protein and mRNA expression, Nox4 promoter activation, and reactive oxygen species production. Mutation of the CC(A/T)6GG box eliminates the synergistic activation of the Nox4 promoter. Jasplakinolide-induced actin polymerization synergizes with TGFβ to facilitate MRTF-dependent Nox4 mRNA expression/promoter activation. Moreover, MRTF inhibition prevents Nox4 expression during TGFβ-induced fibroblast-myofibroblast transition as well. Although necessary, MRTF is insufficient; Nox4 expression also requires TGFβ-activated Smad3 and TAZ/YAP, two contact- and cytoskeleton-regulated Smad3-interacting coactivators. Down-regulation/inhibition of TAZ/YAP mitigates injury-induced epithelial Nox4 expression in vitro and in vivo. These findings uncover new MRTF- and TAZ/YAP-dependent mechanisms, which link cytoskeleton remodeling and redox state and impact epithelial plasticity and myofibroblast transition.
AB - TGFβ-induced expression of the NADPH oxidase Nox4 is essential for fibroblast-myofibroblast transition. Rho has been implicated in Nox4 regulation, but the underlying mechanisms are largely unknown. Myocardin-related transcription factor (MRTF), a Rho/actin polymerization-controlled coactivator of serum response factor, drives myofibroblast transition from various precursors. We have shown that TGFβ is necessary but insufficient for epithelial-myofibroblast transition in intact epithelia; the other prerequisite is the uncoupling of intercellular contacts, which induces Rho-dependent nuclear translocation of MRTF. Because the Nox4 promoter harbors a serum response factor/MRTF cis-element (CC(A/T)6GG box), we asked if MRTF (and thus cytoskeleton organization) could regulate Nox4 expression. We show that Nox4 protein is robustly induced in kidney tubular cells exclusively by combined application of contact uncoupling and TGFβ. Nox4 knockdown abrogates epithelial-myofibroblast transition-associated reactive oxygen species production. Laser capture microdissection reveals increased Nox4 expression in the tubular epithelium also during obstructive nephropathy. MRTF down-regulation/inhibition suppresses TGFβ/contact disruption-provoked Nox4 protein and mRNA expression, Nox4 promoter activation, and reactive oxygen species production. Mutation of the CC(A/T)6GG box eliminates the synergistic activation of the Nox4 promoter. Jasplakinolide-induced actin polymerization synergizes with TGFβ to facilitate MRTF-dependent Nox4 mRNA expression/promoter activation. Moreover, MRTF inhibition prevents Nox4 expression during TGFβ-induced fibroblast-myofibroblast transition as well. Although necessary, MRTF is insufficient; Nox4 expression also requires TGFβ-activated Smad3 and TAZ/YAP, two contact- and cytoskeleton-regulated Smad3-interacting coactivators. Down-regulation/inhibition of TAZ/YAP mitigates injury-induced epithelial Nox4 expression in vitro and in vivo. These findings uncover new MRTF- and TAZ/YAP-dependent mechanisms, which link cytoskeleton remodeling and redox state and impact epithelial plasticity and myofibroblast transition.
KW - Actins
KW - Animals
KW - Cytoskeleton
KW - Epithelium
KW - Fibrosis
KW - Gene Expression Regulation, Enzymologic
KW - Kidney Tubules
KW - LLC-PK1 Cells
KW - Male
KW - Mesoderm
KW - Mice, Inbred C57BL
KW - Muscle Development
KW - Myofibroblasts
KW - NADPH Oxidase
KW - Oxidation-Reduction
KW - Polymerization
KW - Promoter Regions, Genetic
KW - RNA, Messenger
KW - Rats
KW - Reactive Oxygen Species
KW - Swine
KW - Transcription Factors
KW - Up-Regulation
U2 - 10.1074/jbc.M115.674606
DO - 10.1074/jbc.M115.674606
M3 - Journal article
C2 - 26555261
VL - 291
SP - 227
EP - 243
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 1
ER -
ID: 161666599