Association of Sp3 and estrogen receptor alpha with the transcriptionally active trefoil factor 1 promoter in MCF-7 breast cancer cells.
To further explore the role of Sp1 and Sp3 in the estrogen regulated TFF1 gene transcription, chromatin immunoprecipitation (ChIP) assay was used to determine the association of estrogen receptor alpha (ERalpha), Sp1 and Sp3 with the endogenous trefoil factor 1 (TFF1) gene promoter in MCF-7 breast cancer cells. ERalpha and serine 5 phosphorylated RNA polymerase II, the form of RNA polymerase II associated with transcription initiation, were recruited to the TFF1 gene promoter following estrogen addition to MCF-7 cells cultured under estrogen deplete conditions. Both Sp1 and Sp3 were bound to the TFF1 gene promoter before and after estrogen treatment. Using the re-ChIP assay, we demonstrate that either Sp1 or Sp3 but not both bind to a TFF1 promoter. The co-occupancy of ERalpha and Sp1 on TFF1 promoter remains at similar level with and without estrogen, while that of ERalpha and Sp3 increased in the presence of estrogen. Further, we observed increased co-occupancy of Sp3 and serine 5 phosphorylated RNA polymerase II on the TFF1 promoter after estrogen treatment of cells. Taken together, these results provide evidence that Sp3 and ERalpha are involved in the estrogen induced transcription of the TFF1 gene.
Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Manitoba, Canada.
Lin
L
Li
James R
JR
Davie
D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D005260:Female; D006801:Humans; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D051705:Sp3 Transcription Factor / Q000378:metabolism*; D014158:Transcription, Genetic; D000071163:Trefoil Factor-1; D025521:Tumor Suppressor Proteins / Q000235:genetics*
C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha; D016329:Sp1 Transcription Factor; C051838:TFF1 protein, human; D000071163:Trefoil Factor-1; D025521:Tumor Suppressor Proteins; D051705:Sp3 Transcription Factor