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.
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. Developmental Dynamics 238:1574-1581, 2009. (c) 2009 Wiley-Liss, Inc.
Park
Jae-Sun
JS
Division of Intractable Diseases, Center for Biomedical Sciences, Korea National Institute of Health, Seoul, Korea.
Kim
Hyung-Seok
HS
Kim
Jun-Dae
JD
Seo
Jungwon
J
Chung
Kyung-Sook
KS
Lee
Ho-Sa
HS
Huh
Tae-Lin
TL
Jo
Inho
I
Kim
Yong-Ou
YO
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D050980:GATA Transcription Factors / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D006352:Heart Ventricles / Q000196:embryology / Q000378:metabolism; D008969:Molecular Sequence Data; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish / Q000033:anatomy & histology / Q000196:embryology / Q000235:genetics; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
D050980:GATA Transcription Factors; C525957:Gata4 protein, zebrafish; C525956:Gata6 protein, zebrafish; D016376:Oligonucleotides, Antisense; D011993:Recombinant Fusion Proteins; D029961:Zebrafish Proteins; D018995:Myosin Heavy Chains