General method for isolation of DNA sequences that interact with specific nuclear proteins in chromosomes: binding of the high mobility group protein HMG-T to a subset of the protamine gene family

A general method is described for the isolation of the DNA with which specific nuclear proteins interact in chromosomes. This method is based on the covalent photo-cross-linking of nuclear proteins to the DNA sequences, to which they normally bind, by means of irradiation with UV light and the selec...

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Bibliographic Details
Published in:Biochemistry (Easton) 1985-12, Vol.24 (27), p.8021-8028
Main Authors: Blanco, J, States, J. C, Dixon, Gordon H
Format: Article
Language:English
Subjects:
Animals
Antibodies
Base Sequence
Biological and medical sciences
Cell Nucleus - metabolism
Chromatin. Chromosome
Chromatography, Affinity
Chromosomes - metabolism
DNA - isolation & purification
DNA - metabolism
Fundamental and applied biological sciences. Psychology
Genes
High Mobility Group Proteins - metabolism
Liver - metabolism
Molecular and cellular biology
Molecular genetics
Nucleic Acid Hybridization
Nucleoproteins - metabolism
Protamines - genetics
Trout
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Summary:A general method is described for the isolation of the DNA with which specific nuclear proteins interact in chromosomes. This method is based on the covalent photo-cross-linking of nuclear proteins to the DNA sequences, to which they normally bind, by means of irradiation with UV light and the selective retrieval of specific subsets of protein-DNA adducts by using specific antibodies. The application of this procedure to isolate the DNA sequences with which the trout high mobility group protein (HMG-T) interacts has shown that in trout liver this protein associates specifically with DNA sequences in proximity to a subset of the family of protamine genes but not with the histone or vitellogenin genes. From these observations, it appears that the HMG-T protein may be associated with inactive gene sequences.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi00348a028