Regulation of the human transforming growth factor beta type II receptor gene promoter by novel Sp1 sites.
The transforming growth factor-beta (TGF beta) type II receptor (T beta R-II) is responsible for transducing the growth inhibitory signals of TGF beta. The T beta R-II gene promoter lacks both a TATA box and a CAAT box near the transcription initiation site, and has been shown to contain binding sequences for several transcription factors (Sp1, AP1, NF-Y, Cut and ERT) which are important for T beta R-II gene promoter activity in vitro. However, it is still not clear which interactions are important for the regulation of T beta R-II gene promoter activity in vivo. Using in vivo genomic DNA footprinting of normal human epithelial cells (HaCaT), we have identified two novel identical and strongly protected sites (ggggctgg) at positions -59 and -102 of the T beta R-II gene promoter. Mutation of either site significantly reduced promoter activity in transient transfections. Protein binding to these sites, as determined by electrophoretic mobility shift assays (EMSA), was specifically competed with consensus Sp1 oligonucleotides. Furthermore, anti-Sp1/3 antibodies produced band shifts when incubated with the T beta R-II -59 and -102 DNA probes. Importantly, Sp1 protein binding was influenced by the presence of an intact NF-Y binding site at position -83. Our data suggests that both Sp1 and NF-Y may play an important role in regulating T beta R-II gene promoter basal activity in vivo.
Jennings
R
R
Molecular Oncology Program, H. Lee Moffitt Cancer Center and Research Institute, Department of Oncology, University of South Florida, Tampa, Florida, FL 33612, USA.
Alsarraj
M
M
Wright
K L
KL
Muñoz-Antonia
T
T
D001665:Binding Sites; D023081:CCAAT-Binding Factor / Q000502:physiology; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology; D006801:Humans; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics*; D020218:Response Elements; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000502:physiology*; D051705:Sp3 Transcription Factor; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology; D015533:Transcriptional Activation; D014162:Transfection
D023081:CCAAT-Binding Factor; D004268:DNA-Binding Proteins; D018125:Receptors, Transforming Growth Factor beta; D016329:Sp1 Transcription Factor; D014157:Transcription Factors; D051705:Sp3 Transcription Factor; D004247:DNA; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II