Potential for H-DNA in the human MUC1 mucin gene promoter.
Similar imperfect purine/pyrimidine mirror repeat (PMR) elements have previously been identified upstream of the human MUC1 mucin and CFTR genes. These elements confer S1 nuclease sensitivity on isolated plasmid DNA at low pH. We now present a detailed characterization of the non-B DNA structure responsible for S1 nuclease sensitivity upstream of the MUC1 gene. A approximately 90-base pair (bp) DNA fragment containing a 32-bp PMR element termed M-PMR3 was subcloned into a recombinant vector. This fragment conferred S1 nuclease sensitivity on the resulting supercoiled plasmid. High resolution mapping of sites reactive to S1 and P1 nucleases demonstrates that cleavage occurs within the M-PMR3 element. High resolution mapping with chemical agents selective for non-B DNA provides evidence that M-PMR3 adopts an H-DNA structure (intramolecular triple helix) in the less common H-y5 isomer at low pH. This result is observed in the presence or absence of Mg2+. Mutation of the native M-PMR3 element to create perfect homopurine/homopyrimidine mirror symmetry alters the preferred folding to the more common H-y3 triplex DNA isomer. These results demonstrate that imperfections in mirror symmetry can alter the relative stabilities of different H-DNA isomers.
Nelson
K L
KL
Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska 68198-6805, USA.
Becker
N A
NA
Pahwa
G S
GS
Hollingsworth
M A
MA
Maher
L J
LJ
D001483:Base Sequence; D002413:Cations, Divalent / Q000494:pharmacology; D004247:DNA / Q000737:chemistry* / Q000187:drug effects; D006801:Humans; D008274:Magnesium / Q000494:pharmacology; D015335:Molecular Probes; D008969:Molecular Sequence Data; D018396:Mucin-1 / Q000235:genetics*; D009077:Mucins / Q000235:genetics*; D009690:Nucleic Acid Conformation; D009711:Nucleotides / Q000737:chemistry; D011401:Promoter Regions, Genetic; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000494:pharmacology
D002413:Cations, Divalent; D015335:Molecular Probes; D018396:Mucin-1; D009077:Mucins; D009711:Nucleotides; C051802:triplex DNA; D004247:DNA; D015719:Single-Strand Specific DNA and RNA Endonucleases; D008274:Magnesium