A novel function of nuclear nonmuscle myosin regulatory light chain in promotion of xanthine oxidase transcription after myocardial ischemia/reperfusion

Yi Shuai Zhang, Bin Liu, Xiu Ju Luo, Jie Jie Zhang, Nian Sheng Li, Qi Lin Ma, Jun Lin Jiang, Yuan Jian Li, Qingjie Li, Jun Peng

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Nuclear myosin regulates gene transcription and this novel function might be modulated through phosphorylation of the myosin regulatory light chain (p-MLC<inf>20</inf>). Nonmuscle MLC<inf>20</inf> (nmMLC<inf>20</inf>) is also present in the nuclei of cardiomyocytes and a potential nmMLC<inf>20</inf> binding sequence has been identified in the promoter of the xanthine oxidase (XO) gene. Thus, we investigated its function in the regulation of XO transcription after myocardial ischemia/reperfusion (IR). In a rat model of myocardial IR and a cardiomyocyte model of hypoxia/reoxygenation (HR) injury, the cardiac or cell injury, myosin light chain kinase (MLCK) content, XO expression and activity, XO-derived products, and level of nuclear p-nmMLC<inf>20</inf> were detected. Coimmunoprecipitation (co-IP), chromatin immunoprecipitation, DNA pull-down, and luciferase reporter gene assays were used to decipher the molecular mechanisms through which nmMLC<inf>20</inf> promotes XO expression. IR or HR treatment dramatically elevated nuclear p-nmMLC<inf>20</inf> level, accompanied by increased XO expression, activity, and products (H<inf>2</inf>O<inf>2</inf> and uric acid), as well as the IR or HR injury; these effects were ameliorated by inhibition of MLCK or knockdown of nmMLC<inf>20</inf>. Our findings from these experiments demonstrated that nuclear p-nmMLC<inf>20</inf> binds to the consensus sequence GTCGCC in the XO gene promoter, interacts with RNA polymerase II and transcription factor IIB to form a transcription preinitiation complex, and hence activates XO gene transcription. These results suggest that nuclear p-nmMLC<inf>20</inf> plays an important role in IR/HR injury by transcriptionally upregulating XO gene expression to increase oxidative stress in myocardium. Our findings demonstrate nuclear nmMLC<inf>20</inf> as a potential new therapeutic target to combat cardiac IR injury.

Original languageEnglish (US)
Pages (from-to)115-128
Number of pages14
JournalFree Radical Biology and Medicine
Volume83
DOIs
StatePublished - Jun 1 2015

Fingerprint

Myocardial Reperfusion
Myosin Light Chains
Xanthine Oxidase
Transcription
Myocardial Ischemia
Genes
Reperfusion
Myosin-Light-Chain Kinase
Ischemia
Wounds and Injuries
Cardiac Myocytes
Transcription Factor TFIIB
Phosphorylation
Oxidative stress
RNA Polymerase II
Chromatin Immunoprecipitation
Consensus Sequence
Myosins
Uric Acid
Reperfusion Injury

Keywords

  • Free radicals
  • Ischemia
  • Nonmuscle myosin regulatory light chain
  • Reperfusion
  • Transcription
  • Xanthine oxidase

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

Cite this

A novel function of nuclear nonmuscle myosin regulatory light chain in promotion of xanthine oxidase transcription after myocardial ischemia/reperfusion. / Zhang, Yi Shuai; Liu, Bin; Luo, Xiu Ju; Zhang, Jie Jie; Li, Nian Sheng; Ma, Qi Lin; Jiang, Jun Lin; Li, Yuan Jian; Li, Qingjie; Peng, Jun.

In: Free Radical Biology and Medicine, Vol. 83, 01.06.2015, p. 115-128.

Research output: Contribution to journalArticle

Zhang, Yi Shuai ; Liu, Bin ; Luo, Xiu Ju ; Zhang, Jie Jie ; Li, Nian Sheng ; Ma, Qi Lin ; Jiang, Jun Lin ; Li, Yuan Jian ; Li, Qingjie ; Peng, Jun. / A novel function of nuclear nonmuscle myosin regulatory light chain in promotion of xanthine oxidase transcription after myocardial ischemia/reperfusion. In: Free Radical Biology and Medicine. 2015 ; Vol. 83. pp. 115-128.
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abstract = "Nuclear myosin regulates gene transcription and this novel function might be modulated through phosphorylation of the myosin regulatory light chain (p-MLC20). Nonmuscle MLC20 (nmMLC20) is also present in the nuclei of cardiomyocytes and a potential nmMLC20 binding sequence has been identified in the promoter of the xanthine oxidase (XO) gene. Thus, we investigated its function in the regulation of XO transcription after myocardial ischemia/reperfusion (IR). In a rat model of myocardial IR and a cardiomyocyte model of hypoxia/reoxygenation (HR) injury, the cardiac or cell injury, myosin light chain kinase (MLCK) content, XO expression and activity, XO-derived products, and level of nuclear p-nmMLC20 were detected. Coimmunoprecipitation (co-IP), chromatin immunoprecipitation, DNA pull-down, and luciferase reporter gene assays were used to decipher the molecular mechanisms through which nmMLC20 promotes XO expression. IR or HR treatment dramatically elevated nuclear p-nmMLC20 level, accompanied by increased XO expression, activity, and products (H2O2 and uric acid), as well as the IR or HR injury; these effects were ameliorated by inhibition of MLCK or knockdown of nmMLC20. Our findings from these experiments demonstrated that nuclear p-nmMLC20 binds to the consensus sequence GTCGCC in the XO gene promoter, interacts with RNA polymerase II and transcription factor IIB to form a transcription preinitiation complex, and hence activates XO gene transcription. These results suggest that nuclear p-nmMLC20 plays an important role in IR/HR injury by transcriptionally upregulating XO gene expression to increase oxidative stress in myocardium. Our findings demonstrate nuclear nmMLC20 as a potential new therapeutic target to combat cardiac IR injury.",
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AU - Zhang, Jie Jie

AU - Li, Nian Sheng

AU - Ma, Qi Lin

AU - Jiang, Jun Lin

AU - Li, Yuan Jian

AU - Li, Qingjie

AU - Peng, Jun

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