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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1994-12-20
pubmed:abstractText
We have expressed and purified a series of recombinant zinc finger polypeptides derived from the cDNA for the Xenopus 5 S gene-specific transcription factor TFIIIA. Dissociation constants for the interaction of each of the truncated polypeptides with the 5 S gene promoter have been measured using gel mobility shift assays. DNase I footprinting and proteolysis experiments provide additional insights into the interactions of individual fingers within complexes of the truncated proteins. These results are discussed in terms of recently proposed models for the TFIIIA-DNA interaction. The effects of mutations in two of the strongly binding proteins, zf1-3 and zf1-7, on DNA binding affinity have been investigated. Mutations have been made both in putative DNA-contact residues and in the linker regions between zinc fingers. The observed decreases in binding affinity cannot be explained simply in terms of loss of protein-DNA contacts. Our results support a model in which DNA binding is accomplished through sets of interacting zinc fingers that make different energetic contributions to the overall binding of the protein and different contacts with the DNA.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0022-2836
pubmed:author
pubmed:issnType
Print
pubmed:day
18
pubmed:volume
244
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
23-35
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:7966319-Amino Acid Sequence, pubmed-meshheading:7966319-Animals, pubmed-meshheading:7966319-Base Sequence, pubmed-meshheading:7966319-Binding Sites, pubmed-meshheading:7966319-DNA, Ribosomal, pubmed-meshheading:7966319-DNA-Binding Proteins, pubmed-meshheading:7966319-Models, Genetic, pubmed-meshheading:7966319-Molecular Sequence Data, pubmed-meshheading:7966319-Peptide Fragments, pubmed-meshheading:7966319-Promoter Regions, Genetic, pubmed-meshheading:7966319-Protein Binding, pubmed-meshheading:7966319-RNA, Ribosomal, 5S, pubmed-meshheading:7966319-Recombinant Proteins, pubmed-meshheading:7966319-Thermodynamics, pubmed-meshheading:7966319-Transcription Factor TFIIIA, pubmed-meshheading:7966319-Transcription Factors, pubmed-meshheading:7966319-Xenopus, pubmed-meshheading:7966319-Zinc Fingers
pubmed:year
1994
pubmed:articleTitle
Relative contributions of the zinc fingers of transcription factor IIIA to the energetics of DNA binding.
pubmed:affiliation
Department of Molecular Biology, Scripps Research Institute, La Jolla, CA 92037.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't