TY - JOUR
T1 - Stimulation of the murine type II transforming growth factor-β receptor promoter by the transcription factor Egr-1
AU - Wilder, Phillip J.
AU - Bernadt, Cory T.
AU - Kim, Jae Hwan
AU - Rizzino, Angie
PY - 2002/11/1
Y1 - 2002/11/1
N2 - Previous studies have demonstrated that differentiation of murine embryonal carcinoma (EC) cells leads to the appearance of high affinity receptors for transforming growth factor-β (TGF-β). Subsequently, it was demonstrated that differentiation of F9 EC cells leads to increases in the transcription of the type II TGF-β-receptor gene (TβR-II) and leads to significant increases in the steady-state levels of TβR-II mRNA. Analysis of the human TβR-II promoter in F9-differentiated cells identified several cis-regulatory elements that influence the activity of the promoter, including a CRE/ATF site and a CCAAT box motif. In the work described in this report, we focused on the effect of the transcription factor Egr-1 on the murine TβR-II promoter. We have identified an Egr-1 response-element ∼150 bp upstream of the major transcription start site of the murine TβR-II gene. We demonstrate by electrophoretic mobility shift analysis (EMSA) that this cis-regulatory element binds Egr-1, and we demonstrate that disruption of this site eliminates the response to Egr-1. As part of this analysis, we also examined the effect of Egr-1 on human TβR-II promoter. In contrast to a previous report, which reported that Egr-1 inhibits expression of human TβR-II promoter/reporter gene constructs, we did not observe an inhibitory effect of Egr-1 that was specific for the human TβR-II promoter. Taken together, the findings described in this report identify important differences between the human and the murine TβR-II promoter, and our findings identify an Egr-1 cis-regulatory element that is capable of stimulating the activity of the murine TβR-II promoter.
AB - Previous studies have demonstrated that differentiation of murine embryonal carcinoma (EC) cells leads to the appearance of high affinity receptors for transforming growth factor-β (TGF-β). Subsequently, it was demonstrated that differentiation of F9 EC cells leads to increases in the transcription of the type II TGF-β-receptor gene (TβR-II) and leads to significant increases in the steady-state levels of TβR-II mRNA. Analysis of the human TβR-II promoter in F9-differentiated cells identified several cis-regulatory elements that influence the activity of the promoter, including a CRE/ATF site and a CCAAT box motif. In the work described in this report, we focused on the effect of the transcription factor Egr-1 on the murine TβR-II promoter. We have identified an Egr-1 response-element ∼150 bp upstream of the major transcription start site of the murine TβR-II gene. We demonstrate by electrophoretic mobility shift analysis (EMSA) that this cis-regulatory element binds Egr-1, and we demonstrate that disruption of this site eliminates the response to Egr-1. As part of this analysis, we also examined the effect of Egr-1 on human TβR-II promoter. In contrast to a previous report, which reported that Egr-1 inhibits expression of human TβR-II promoter/reporter gene constructs, we did not observe an inhibitory effect of Egr-1 that was specific for the human TβR-II promoter. Taken together, the findings described in this report identify important differences between the human and the murine TβR-II promoter, and our findings identify an Egr-1 cis-regulatory element that is capable of stimulating the activity of the murine TβR-II promoter.
KW - Early growth response-1
KW - Embryonal carcinoma cells
KW - Gene regulation
KW - Transforming growth factor-β-receptor
UR - http://www.scopus.com/inward/record.url?scp=0036836544&partnerID=8YFLogxK
U2 - 10.1002/mrd.10165
DO - 10.1002/mrd.10165
M3 - Article
C2 - 12237943
AN - SCOPUS:0036836544
SN - 1040-452X
VL - 63
SP - 282
EP - 290
JO - Molecular Reproduction and Development
JF - Molecular Reproduction and Development
IS - 3
ER -