Triple helix formation with the promoter of human alpha1(I) procollagen gene by an antiparallel triplex-forming oligodeoxyribonucleotide.
The promoters of alpha1(I) procollagen genes of vertebrates contain two contiguous stretches of polypyrimidine/polypurine sequences, referred to as C1 (-140 to -170) and C2 (-171 to -200). Antiparallel triplex-forming upstream oligonucleotides form efficient triplexes with C1. The C1 tract of human differs from rodent alpha1(I) promoters by 7 nt which are mainly A-->G transitions. Human triplex-forming oligodeoxyribonucleotide (TFO) formed stable triplexes efficiently with a K d of approximately 10-20 nM compared with a K d of approximately 100 nM for rodent TFO. Mutational analysis indicated that 3 or 4 nt (-153 to -155) are sufficient for this higher affinity. TFOs specific for human C1 inhibited transcription from human promoter both in vitro in HeLa cell nuclear extracts and in vivo in cultured chick embryo fibroblasts.
Nakanishi
M
M
Department of Molecular Microbiology and Immunology and Division of Cardiology, School of Medicine, University of Missouri-Columbia, Columbia, MO 65212, USA.
Weber
K T
KT
Guntaka
R V
RV
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002642:Chick Embryo; D003001:Cloning, Molecular; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D051379:Mice; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011347:Procollagen / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D014162:Transfection
D009838:Oligodeoxyribonucleotides; D011347:Procollagen