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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
7
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pubmed:dateCreated |
1989-6-14
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pubmed:abstractText |
The equilibrium constant was determined for the binding of the transcription factor Sp1 to a single consensus GC box DNA recognition site, (5'-GGGGCGGGGC-3'). For these experiments, single copies of the recognition site were synthesized and cloned in a standard plasmid background. Binding was measured either by a footprinting assay modified so that the binding reaction was at equilibrium, or by a gel mobility shift assay. The concentration of active Sp1 in the reactions and the dissociation constant were determined by computer-assisted fitting to theoretical curves. Values for the dissociation constant obtained in different experiments ranged from 4.1 X 10(-10) M to 5.3 X 10(-10) M. Several variants of the consensus recognition site were also tested. An A-substituted variant (5'-GGGGAGGGGC-3') and a T-substituted variant (5'-GGGGTGGGGC-3') were bound 3-fold and 6-fold more weakly than the consensus site, respectively. A G-substituted variant (5'-GGGGGGGGGC-3') was bound at least 30-fold more weakly than the consensus site. These findings help distinguish between alternative models for Sp1-DNA recognition. They are consistent with the presence of specific hydrogen-bond contacts between Sp1 and the central C-G base pair, but provide no particular evidence to support a model where local DNA structure is the dominant factor in the interaction.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-1062791,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-1092468,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-113409,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-2842864,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-2992804,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3003068,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3008338,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3056778,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3281160,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3319186,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3351924,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3461465,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3464963,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3529394,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3685975,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3773731,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3796614,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-3945313,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-6095100,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-6187469,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2717405-6313230
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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 |
Apr
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pubmed:issn |
0305-1048
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
11
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pubmed:volume |
17
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
2639-53
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:2717405-Base Composition,
pubmed-meshheading:2717405-Base Sequence,
pubmed-meshheading:2717405-Binding Sites,
pubmed-meshheading:2717405-DNA,
pubmed-meshheading:2717405-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:2717405-Genetic Variation,
pubmed-meshheading:2717405-HeLa Cells,
pubmed-meshheading:2717405-Humans,
pubmed-meshheading:2717405-Kinetics,
pubmed-meshheading:2717405-Nucleotide Mapping,
pubmed-meshheading:2717405-Oligonucleotides,
pubmed-meshheading:2717405-Transcription Factors
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pubmed:year |
1989
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pubmed:articleTitle |
Measurement of the binding of transcription factor Sp1 to a single GC box recognition sequence.
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pubmed:affiliation |
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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