Role for cGATA-5 in transcriptional regulation of the embryonic chicken pepsinogen gene by epithelial-mesenchymal interactions in the developing chicken stomach.
A gene encoding embryonic chicken pepsinogen (ECPg), a zymogen of the digestive enzyme pepsin, is expressed specifically in epithelial cells of glands of embryonic stage proventriculus (glandular stomach) under the influence of mesenchyme. We found four GATA and one Sox binding motifs in 1.1 kb of the 5' flanking region of the ECPg gene which are essential to the organ-specific expression of the gene. The expression of cGATA-5 and cSox2 in the proventriculus from day 6 to day 12 of incubation was therefore analyzed. cGATA-5 was more strongly expressed in glandular epithelial cells than in luminal epithelial cells, while cSox2 gene expression was weaker in glandular epithelial cells. Using heterologous recombination explants we also discovered that the expression of cGATA-5 and cSox2 in epithelial cells was affected by mesenchyme when the latter induced ECPg gene expression in epithelial cells. Introduction of expression constructs into epithelial cells by electroporation demonstrated that cGATA-5 upregulated transcription of a reporter luciferase gene via a cis element in the 5' flanking region of the ECPg gene. The gel mobility shift assay revealed that the cGATA-5 protein specifically binds to the GATA binding sites. cSox2 downregulated the activity of luciferase but it was not through the Sox binding motif. These results suggest that cGATA-5 positively regulates transcription of the ECPg gene and is involved in spatial regulation of the pepsinogen gene during development.
Sakamoto
N
N
Department of Biological Sciences, The Graduate School of Science, Tokyo Metropolitan University, 1-1 Minamiohsawa, Hachioji, Tokyo, 192-0397, Japan.
Fukuda
K
K
Watanuki
K
K
Sakai
D
D
Komano
T
T
Scotting
P J
PJ
Yasugi
S
S
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000378:metabolism* / Q000502:physiology*; D018274:Electroporation; D050992:GATA5 Transcription Factor; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D024002:HMGB Proteins; D017404:In Situ Hybridization, Fluorescence; D007413:Intestinal Mucosa / Q000196:embryology / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D008648:Mesoderm / Q000378:metabolism*; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000378:metabolism; D020116:Pepsinogen A / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011995:Recombination, Genetic; D055748:SOXB1 Transcription Factors; D013270:Stomach / Q000196:embryology*; D014157:Transcription Factors / Q000096:biosynthesis / Q000378:metabolism* / Q000502:physiology*; D014158:Transcription, Genetic; D014162:Transfection
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D050992:GATA5 Transcription Factor; D024002:HMGB Proteins; D009687:Nuclear Proteins; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors; D020116:Pepsinogen A; D008156:Luciferases