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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
10
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pubmed:dateCreated |
1990-10-18
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pubmed:databankReference |
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pubmed:abstractText |
The complete DNA sequence of the 1.7 kilobase pairs (kbp) 5' of the rat growth hormone gene (rGH) has been determined and analyzed for Z-DNA-forming potential. Regions of alternating purine-pyrimidine (APP) sequences located between -1047 and -986 [(GT)31], between -445 and -433 bp, and between -426 and -403 bp relative to the rGH RNA transcription initiation site were identified and shown to form Z-DNA in negatively supercoiled plasmids by two-dimensional gel electrophoresis. Free-energy calculations indicated that Z-DNA forms most readily in the proximal Z-DNA regions. Diethyl pyrocarbonate footprinting of physiologically supercoiled plasmid DNA confirmed the presence of Z-DNA from -444 to -404 bp spanning the two most proximal APP sequences and a short non-APP sequence in between. DNA sequence analysis also predicted a region of DNA curvature near this proximal Z-DNA region. Formation of Z-DNA in the distal Z-DNA region consisting of a (GT)31 repeat was constrained at physiological plasmid superhelical densities. This may be related to the presence of DNA sequences (-1584 to -1559) 512 bp upstream of (GT)31 that undergo cruciform formation and thereby utilize the available free energy. Removal of 580 bp containing the cruciform region resulted in Z-DNA formation within (GT)31, thus demonstrating that deletion mutations can exert topological changes at a distance within the rGH 5'-flanking region. Methylation of two specific cytosines in the rGH 5'-flanking DNA that have been associated with inhibition of rGH promoter activity had no effect on Z-DNA formation. No evidence for DNA secondary structure formation within the rGH second exon-intron or 3'-flanking region was observed. We conclude that the rGH 5'-flanking region undergoes secondary-structure formation at physiological superhelical densities, thus providing a potential mechanism(s) for modulating rGH activity.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-174079,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-2158081,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-2544228,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-2689449,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-2780286,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-3284886,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-3516005,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-3608986,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-3718951,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-3774545,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/2398895-7357619
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0270-7306
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
10
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
5378-87
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:2398895-Alkylation,
pubmed-meshheading:2398895-Animals,
pubmed-meshheading:2398895-Base Sequence,
pubmed-meshheading:2398895-Cloning, Molecular,
pubmed-meshheading:2398895-DNA,
pubmed-meshheading:2398895-Electrophoresis, Gel, Two-Dimensional,
pubmed-meshheading:2398895-Growth Hormone,
pubmed-meshheading:2398895-Hydrogen Bonding,
pubmed-meshheading:2398895-Methylation,
pubmed-meshheading:2398895-Molecular Sequence Data,
pubmed-meshheading:2398895-Nucleic Acid Conformation,
pubmed-meshheading:2398895-Plasmids,
pubmed-meshheading:2398895-Promoter Regions, Genetic,
pubmed-meshheading:2398895-Rats,
pubmed-meshheading:2398895-Restriction Mapping,
pubmed-meshheading:2398895-Thermodynamics
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pubmed:year |
1990
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pubmed:articleTitle |
Z-DNA formation in the rat growth hormone gene promoter region.
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