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Nuclear cardiac myosin light chain 2 modulates NADPH oxidase 2 expression in myocardium: a novel function beyond muscle contraction

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Abstract

Recent studies demonstrated that NADPH oxidase 2 (NOX2) expression in myocardium after ischemia–reperfusion (IR) is significantly upregulated. However, the underlying mechanisms remain unknown. This study aims to determine if nuclear cardiac myosin light chain 2 (MYL2), a well-known regulatory subunit of myosin, functions as a transcription factor to promote NOX2 expression following myocardial IR in a phosphorylation-dependent manner. We examined the phosphorylation status of nuclear MYL2 (p-MYL2) in a rat model of myocardial IR (left main coronary artery subjected to 1 h ligation and 3 h reperfusion) injury, which showed IR injury and upregulated NOX2 expression as expected, accompanied by elevated H2O2 and nuclear p-MYL2 levels; these effects were attenuated by inhibition of myosin light chain kinase (MLCK). Next, we explored the functional relationship of nuclear p-MYL2 with NOX2 expression in H9c2 cell model of hypoxia-reoxygenation (HR) injury. In agreement with our in vivo findings, HR treatment increased apoptosis, NOX2 expression, nuclear p-MYL2 and H2O2 levels, and the increases were ameliorated by inhibition of MLCK or knockdown of MYL2. Finally, molecular biology techniques including co-immunoprecipitation (Co-IP), chromatin immunoprecipitation (ChIP), DNA pull-down and luciferase reporter gene assay were utilized to decipher the molecular mechanisms. We found that nuclear p-MYL2 binds to the consensus sequence AGCTCC in NOX2 gene promoter, interacts with RNA polymerase II and transcription factor IIB to form a transcription preinitiation complex, and thus activates NOX2 gene transcription. Our results demonstrate that nuclear MYL2 plays an important role in IR injury by transcriptionally upregulating NOX2 expression to enhance oxidative stress in a phosphorylation-dependent manner.

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Acknowledgments

This work was supported by the Major Research Plan of the National Natural Science Foundation of China (No. 91439104 to Jun Peng), National Nature Science Foundation of China (No. 81373409 to Jun Peng; No.81370250 to Qi-Lin Ma), Hunan Provincial Natural Science Foundation of China (No.13JJ2008 to Jun Peng) and Doctoral Fund of Ministry of Education of China (No. 20120162110056 to Jun Peng).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Pharmacology, School of Pharmaceutical Sciences, Central South University, Changsha, 410078, China

    Yi-Shuai Zhang, Bin Liu, Xiu-Ju Luo, Ting-Bo Li, Jie-Jie Zhang, Jing-Jie Peng, Xiao-Jie Zhang, Chang-Ping Hu, Yuan-Jian Li & Jun Peng

  2. Hunan Provincial Key Laboratory of Cardiovascular Research, School of Pharmaceutical Sciences, Central South University, Changsha, 410078, China

    Yi-Shuai Zhang, Ting-Bo Li, Jie-Jie Zhang, Xiao-Jie Zhang, Chang-Ping Hu, Yuan-Jian Li & Jun Peng

  3. Department of Cardiovascular Medicine, Xiangya Hospital, Central South University, Changsha, 410008, China

    Qi-Lin Ma

  4. Department of Internal Medicine, The University of Texas Medical Branch at Galveston, Galveston, TX, 77555-1083, USA

    Qingjie Li

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  1. Yi-Shuai Zhang
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Correspondence to Jun Peng.

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Zhang, YS., Liu, B., Luo, XJ. et al. Nuclear cardiac myosin light chain 2 modulates NADPH oxidase 2 expression in myocardium: a novel function beyond muscle contraction. Basic Res Cardiol 110, 38 (2015). https://doi.org/10.1007/s00395-015-0494-5

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  • DOI: https://doi.org/10.1007/s00395-015-0494-5

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