title: NFAT5-mediated CACNA1C expression is critical for cardiac electrophysiological development and maturation.
abstract: Entry of calcium into cardiomyocyte via L-type calcium channel (LTCC) is fundamental to cardiac contraction. CACNA1C, a type of LTCC and a hallmark of a matured ventricular myocyte, is developmentally regulated. Here, we identified 138 potential transcription factors by a comparative genomic study on 5-kb promoter regions of CACNA1C gene across eight vertebrate species, and showed that six factors were developmentally regulated with the expression of Cacna1c in mouse P19cl6 in vitro cardiomyocyte differentiation model. We further demonstrated that the nuclear factor of activated T cells 5 (Nfat5) bound to a consensus sequence TGGAAGCGTTC and activated the transcription of Cacna1c. The siRNA-mediated knockdown of Nfat5 suppressed the expression of Cacna1c and decreased L-type calcium current in mouse neonatal cardiomyocytes. Furthermore, morpholino-mediated knockdown of nfat5 in zebrafish prohibited the expression of cacna1c and resulted in a non-contractile ventricle, while over-expression of either cacna1c or nfat5 rescued this impaired phenotype. Thus, NFAT5-mediated expression of CACNA1C is evolutionarily conserved and critical for cardiac electrophysiological development and maturation of cardiomyocyte.
  
  
  
  Nfat5 binds to a consensus sequence TGGAAGCGTTC in the promoter of Cacna1c. Nfat5 activates the transcription of Cacna1c. Nfat5 knockdown suppresses Cacna1c expression, decreases L-type calcium current, and results in non-beating ventricle. NFAT5-mediated expression of CACNA1C is evolutionarily conserved. NFAT5-mediated CACNA1C expression is critical for cardiac electrophysiological development and maturation.
authors.affiliation: Department of Human Population Genetics, Institute of Molecular Medicine, Peking University, 5 Yiheyuan Road, Beijing, 100871, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, 5 Yiheyuan Road, Beijing, 100871, China.
authors.forename: Nai-Zhong
authors.initials: NZ
authors.lastname: Zheng
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing, 100101, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Yuan
authors.affiliation: Department of Human Population Genetics, Institute of Molecular Medicine, Peking University, 5 Yiheyuan Road, Beijing, 100871, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Xu
authors.affiliation: Department of Human Population Genetics, Institute of Molecular Medicine, Peking University, 5 Yiheyuan Road, Beijing, 100871, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing, 100101, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Gu
authors.affiliation: Department of Human Population Genetics, Institute of Molecular Medicine, Peking University, 5 Yiheyuan Road, Beijing, 100871, China.
authors.forename: Gu-Yan
authors.initials: GY
authors.lastname: Zheng
authors.affiliation: School of Electronics Engineering and Computer Science, Peking University, 5 Yiheyuan Road, Beijing, 100871, China.
authors.forename: Guo-Jie
authors.initials: GJ
authors.lastname: Luo
authors.affiliation: Department of Biomedical Engineering, Lerner Research Institute, The Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH, 44195, USA.
authors.forename: Chun
authors.initials: C
authors.lastname: Fan
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing, 100101, China.
authors.forename: Guang-Ju
authors.initials: GJ
authors.lastname: Ji
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, 5 Yiheyuan Road, Beijing, 100871, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Department of Human Population Genetics, Institute of Molecular Medicine, Peking University, 5 Yiheyuan Road, Beijing, 100871, China. caohuiqing@pku.edu.cn.
authors.forename: Huiqing
authors.initials: H
authors.lastname: Cao
authors.affiliation: Department of Human Population Genetics, Institute of Molecular Medicine, Peking University, 5 Yiheyuan Road, Beijing, 100871, China. tianxiaoli@pku.edu.cn.
authors.forename: Xiao-Li
authors.identifier: 0000-0002-0868-7025
authors.initials: XL
authors.lastname: Tian
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D015415:Biomarkers; D020746:Calcium Channels, L-Type / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D002478:Cells, Cultured; D016384:Consensus Sequence; D055724:Electrophysiological Phenomena / Q000235:genetics; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006801:Humans; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051381:Rats; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish
chemical_list: D015415:Biomarkers; C504728:Cacna1c protein, rat; D020746:Calcium Channels, L-Type; C118125:NFAT5 protein, human; D014157:Transcription Factors
keywords: Cardiac development; Comparative genomic; L-type calcium channel; NFAT5; Transcriptional factor