ZFIN ID: ZDB-PUB-090518-20
Isolation of a ventricle-specific promoter for the zebrafish ventricular myosin heavy chain (vmhc) gene and its regulation by GATA factors during embryonic heart development
Park, J.S., Kim, H.S., Kim, J.D., Seo, J., Chung, K.S., Lee, H.S., Huh, T.L., Jo, I., and Kim, Y.O.
Date: 2009
Source: Developmental dynamics : an official publication of the American Association of Anatomists   238(6): 1574-1581 (Journal)
Registered Authors: Huh, Tae-Lin, Kim, Hyung-Seok, Kim, Jun-Dae
Keywords: Myh-15, β-MHC, GATA4, GATA6, transgenic, cardiac, myocardium
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Base Sequence
  • GATA Transcription Factors/genetics
  • GATA Transcription Factors/metabolism*
  • Gene Expression Regulation, Developmental*
  • Genes, Reporter
  • Heart Ventricles*/embryology
  • Heart Ventricles*/metabolism
  • Molecular Sequence Data
  • Myosin Heavy Chains*/genetics
  • Myosin Heavy Chains*/metabolism
  • Oligonucleotides, Antisense/genetics
  • Oligonucleotides, Antisense/metabolism
  • Promoter Regions, Genetic*
  • Recombinant Fusion Proteins/genetics
  • Recombinant Fusion Proteins/metabolism
  • Zebrafish*/anatomy & histology
  • Zebrafish*/embryology
  • Zebrafish*/genetics
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
PubMed: 19441085 Full text @ Dev. Dyn.
FIGURES
ABSTRACT
We investigated chamber-specific gene expression by isolating a 2.2-kb polymerase chain reaction product containing the 5'-flanking region of the zebrafish ventricular myosin heavy-chain gene (vmhc). Promoter analysis revealed that the fragment, consisting of nucleotides from -301 to +26, is sufficient for vmhc promoter activity. Among several putative cis-acting elements in the region, a GATA-binding site was identified to be crucial for the activity of the promoter, as evidenced by the complete abolishment of promoter activity by a single nucleotide substitution of GATA-binding site (-287, C-->T). Knockdown of GATA-binding proteins 4 and 6 (GATA4 and -6) by their antisense morpholino oligonucleotides resulted in reduced green fluorescent protein (GFP) reporter gene and endogenous vmhc expression. These findings suggest that GATA4 and -6 play a key role in the regulation of vmhc expression in the ventricle. In addition, the vmhc promoter and the transgenic zebrafish (vmhc:gfp) are useful tools to study the formation and function of the ventricle.
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