NDLI: Specific photocrosslinking of DNA-protein complexes: identification of contacts between integration host factor and its target DNA.

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Specific photocrosslinking of DNA-protein complexes: identification of contacts between integration host factor and its target DNA.

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Specific photocrosslinking of DNA-protein complexes: identification of contacts between integration host factor and its target DNA.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Yang, S. W. Nash, H. A.
Description	Author Affiliation: Yang SW ( Laboratory of Molecular Biology, National Institute of Mental Health, Bethesda, MD 20892-4034.);
Abstract	Azide moieties have been specifically placed in the backbone of DNA by chemical coupling between azidophenacyl bromide and uniquely positioned phosphorothioate residues. The derivatized DNA forms specific complexes with a DNA-binding protein and, following irradiation with 302-nm light, makes specific crosslinks to the protein. Isolation of this covalent complex, followed by tryptic digestion and Edman degradation of the resulting crosslinked peptide, identifies the portion of the protein that is near the derivatized segment of the target DNA. We use this method to probe the interaction between a specific DNA sequence and integration host factor (IHF) protein. A single IHF heterodimer is known to contact > 25 bp of DNA and thereby introduce a sharp bend. Two segments of a typical IHF site were derivatized with aryl azide. Although the segments were separated by only 5 bp, they crosslinked to different subunits of IHF. The locations of the crosslinks support our current view for the way IHF protein binds to and bends its specific targets.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	25
Volume Number	91
Language	English
Publisher	National Academy of Sciences
Publisher Date	1994-01-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Bacterial Proteins Chemistry Cross-Linking Reagents DNA-Binding Proteins DNA Nucleic Acid Conformation Oligodeoxyribonucleotides Protein Structure, Secondary Amino Acid Sequence Azides Isolation & Purification Metabolism Binding Sites Integration Host Factors Models, Molecular Molecular Sequence Data Peptide Fragments Thionucleotides Trypsin Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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