RRC ID |
42951
|
Author |
Susa T, Nakayama M, Kitahara K, Kimoto F, Kato T, Kato Y.
|
Title |
Homeodomain transcription factor Hesx1/Rpx occupies Prop-1 activation sites in porcine follicle stimulating hormone (FSH) beta subunit promoter.
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Journal |
Biochem Biophys Res Commun
|
Abstract |
Homeodomain repressor factor Hesx1/Rpx plays a crucial role in the formation of Rathke's pouch at the start of pituitary organogenesis and represses the Prop-1-dependent expression of Pit-1 gene, which promotes the differentiation of Pit-1-dependent hormone producing cells. Recently, we discovered a novel function of Prop-1 by which it activates the porcine follicle stimulating hormone beta subunit (FSHbeta) gene through Fd2 region (-852/-746). The present study aimed to determine whether Hesx1 exerts its role in the Prop-1-dependent activation of FSHbeta gene. Transient transfection assay for the porcine FSHbeta promoter -985/+10, electrophoretic mobility shift assay (EMSA) and DNase I footprinting analysis for Fd2 region were carried out. Transfection assay in GH3 cells demonstrated that expression of Hesx1 alone does not change the promoter activity but the coexpression with Prop-1 represses the Prop-1-dependent activation of FSHbeta promoter. Similar results were obtained for the mutant reporter vector deleting the region -745/-104 indicating that Fd2 region is a target site of Hesx1 as well as Prop-1. EMSA and DNase I footprinting analysis using recombinant Hesx1 and Prop-1 protein demonstrated that Hesx1 and Prop-1 certainly bind to the AT-rich regions in a different manner. These results suggest that Hesx1 blocks the advanced expression of FSHbeta gene in the early stage of pituitary development, and Prop-1 thereafter appears and activates this gene.
|
Volume |
357(3)
|
Pages |
712-7
|
Published |
2007-6-8
|
DOI |
10.1016/j.bbrc.2007.03.197
|
PII |
S0006-291X(07)00702-4
|
PMID |
17445765
|
MeSH |
Animals
Base Sequence
Binding Sites / genetics
Cell Line, Tumor
DNA / genetics
DNA / metabolism
DNA Footprinting / methods
Deoxyribonuclease I / metabolism
Electrophoretic Mobility Shift Assay
Follicle Stimulating Hormone, beta Subunit / genetics*
Gene Expression Regulation
Genetic Vectors / genetics
Homeodomain Proteins / genetics
Homeodomain Proteins / metabolism*
Mice
Molecular Sequence Data
Promoter Regions, Genetic / genetics*
Protein Binding
Repressor Proteins
Swine
Transfection
|
IF |
2.985
|
Times Cited |
6
|
WOS Category
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
Resource |
Human and Animal Cells |
|