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PMID: 1569931 Published · ppublish English Journal Article

Only two of the five zinc fingers of the eukaryotic transcriptional repressor PRDI-BF1 are required for sequence-specific DNA binding.

Molecular and cellular biology ·Vol. 12 ·No. 5 ·1992-05-00 ·Pages 1940-9

Keller AD, Maniatis T

Abstract

The eukaryotic transcriptional repressor PRDI-BF1 contains five zinc fingers of the C2H2 type, and the protein binds specifically to PRDI, a 14-bp regulatory element of the beta interferon gene promoter. We have investigated the amino acid sequence requirements for specific binding to PRDI and found that the five zinc fingers and a short stretch of amino acids N terminal to the first finger are necessary and sufficient for PRDI-specific binding. The contribution of individual zinc fingers to DNA binding was investigated by inserting them in various combinations into another zinc finger-containing DNA-binding protein whose own fingers had been removed. We found that insertion of PRDI-BF1 zinc fingers 1 and 2 confer PRDI-binding activity on the recipient protein. In contrast, the insertion of PRDI-BF1 zinc fingers 2 through 5, the insertion of zinc finger 1 or 2 alone, and the insertion of zinc fingers 1 and 2 in reverse order did not confer PRDI-binding activity. We conclude that the first two PRDI-BF1 zinc fingers together are sufficient for the sequence-specific recognition of PRDI.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Binding Sites Chromosome Deletion Deoxyribonuclease I Genetic Vectors Interferon-beta/genetics Introns Molecular Sequence Data Mutagenesis, Site-Directed Oligonucleotide Probes Promoter Regions, Genetic RNA Splicing Regulatory Sequences, Nucleic Acid Repressor Proteins/genetics,metabolism Restriction Mapping Transcription Factors Transcription, Genetic Zinc Fingers/genetics
Chemicals
Oligonucleotide Probes Repressor Proteins Transcription Factors Interferon-beta Deoxyribonuclease I
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Keller A D
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, Massachusetts 02138.
Maniatis T
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48 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.5.1940-1949.1992
Published
1992-05-00
Pages
1940-9
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC364364
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.12.5.1940-1949.1992 PMC Full Text View journal details
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