TY - JOUR
T1 - Evidence for three major transcription activation elements in the proximal mouse proα2(I) collagen promoter
AU - Hasegawa, Tadao
AU - Zhou, Xin
AU - Garrett, Lee Ann
AU - Cristy Ruteshouser, E.
AU - Maity, Sankar N.
AU - De Crombrugghe, Benoit
N1 - Funding Information:
We thank Sandra McKinney and Heidi Eberspaecher for excellent technical assistance and Patricia McCauley for editorial assistance. This work was supported by National Institute of Health grant HL41264 (to B. de C.) and the Japan Foundation for Aging and Health (to T. H.).
PY - 1996
Y1 - 1996
N2 - In vivo transient expression and in vitro transcription experiments indicated that a segment between -170 and -40 bp upstream of the start of transcription of the mouse proα2(I) collagen gene was essential to activate transcription. DNase I protection experiments identified three strong footprints in this segment. Experiments with deletion mutants encompassing the sequences defined by these three footprints indicated that each of the three elements contributed to the transcriptional activity of the promoter. All three elements are GC-rich, redundant sites for a complex set of DNA binding proteins that includes SP1, other proteins that bind to an SP1 consensus site and proteins that bind to a Krox consensus site. In addition, the segment corresponding to the most proximal footprint also binds the multimeric CCAAT binding protein CBF. Addition of an excess amount of oligonucleotides corresponding to either of the two distal footprints significantly inhibited in vitro transcription of the -350 bp proα2(I) collagen promoter. Anti-SP1 antibodies that completely inhibited transcription of the early SV40 promoter had little effect on transcription of the wild-type -350 bp promoter, suggesting that SP1 has only a minor role in activity of this promoter. Our results show that the segment between base pairs -170 and -40 of the proα2(I) collagen promoter, which contains redundant binding sites for a complex set of nuclear proteins, is essential in the transcriptional activity of this promoter in fibroblasts.
AB - In vivo transient expression and in vitro transcription experiments indicated that a segment between -170 and -40 bp upstream of the start of transcription of the mouse proα2(I) collagen gene was essential to activate transcription. DNase I protection experiments identified three strong footprints in this segment. Experiments with deletion mutants encompassing the sequences defined by these three footprints indicated that each of the three elements contributed to the transcriptional activity of the promoter. All three elements are GC-rich, redundant sites for a complex set of DNA binding proteins that includes SP1, other proteins that bind to an SP1 consensus site and proteins that bind to a Krox consensus site. In addition, the segment corresponding to the most proximal footprint also binds the multimeric CCAAT binding protein CBF. Addition of an excess amount of oligonucleotides corresponding to either of the two distal footprints significantly inhibited in vitro transcription of the -350 bp proα2(I) collagen promoter. Anti-SP1 antibodies that completely inhibited transcription of the early SV40 promoter had little effect on transcription of the wild-type -350 bp promoter, suggesting that SP1 has only a minor role in activity of this promoter. Our results show that the segment between base pairs -170 and -40 of the proα2(I) collagen promoter, which contains redundant binding sites for a complex set of nuclear proteins, is essential in the transcriptional activity of this promoter in fibroblasts.
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M3 - Article
C2 - 8774909
AN - SCOPUS:0029779955
SN - 0305-1048
VL - 24
SP - 3253
EP - 3260
JO - Nucleic acids research
JF - Nucleic acids research
IS - 16
ER -