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PMID: 3039460 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Site-specific DNA binding of nuclear factor I: effect of the spacer region.

Nucleic acids research ·Vol. 15 ·No. 14 ·1987-07-24 ·Pages 5545-59

Gronostajski RM

Abstract

Nuclear factor I (NFI) is a site-specific DNA binding protein required for the replication of adenovirus type 2 DNA in vitro and in vivo. To study sequence requirements for the interaction of NFI with DNA, we have measured the binding of the protein to a variety of synthetic sites. Binding sites for NFI (FIB sites) were previously shown to contain a consensus sequence composed of 2 motifs, TGG (Motif 1), and GCCAA (Motif 2), separated by a 6 or 7bp spacer region. To assess conserved sequences in the spacer region and flanking sequences which affect NFI binding, we have isolated clones from oligonucleotide libraries that contain the two motifs flanked by 3 degenerate nucleotides and separated by degenerate spacer regions of 6 or 7 nucleotides. With a 6bp spacer region, a strong bias exists for a C or A residue in the first position of the spacer. Sites with a 7bp spacer region contain a G and C or A residue at the first and second positions, respectively, of the spacer, but also possess conserved residues at other positions of the site.

MeSH Terms
Base Sequence CCAAT-Enhancer-Binding Proteins Cell Nucleus/metabolism Cloning, Molecular DNA/metabolism DNA Restriction Enzymes DNA-Binding Proteins/metabolism HeLa Cells/metabolism Humans NFI Transcription Factors Nuclear Proteins Oligodeoxyribonucleotides/metabolism Structure-Activity Relationship Transcription Factors Y-Box-Binding Protein 1
Chemicals
CCAAT-Enhancer-Binding Proteins DNA-Binding Proteins NFI Transcription Factors Nuclear Proteins Oligodeoxyribonucleotides Transcription Factors Y-Box-Binding Protein 1 YBX1 protein, human DNA DNA Restriction Enzymes
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Gronostajski R M
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47 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.14.5545
Published
1987-07-24
Pages
5545-59
Language
English
Region
England
NLM ID
0411011
PMCID
PMC306006
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/15.14.5545 PMC Full Text View journal details
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