A method for mapping intranuclear protein-DNA interactions and its application to a nuclease hypersensitive site.

Abstract:

:We have devised a method for mapping sites on DNA within the nucleus that are protected against nuclease attack by interaction with bound protein or other factors. This "footprinting" method uses an end-labeled sequence-specific DNA probe, which is annealed to the DNA from nuclear digests under carefully controlled conditions. The annealed complexes are treated with single-strand-specific nuclease, the resulting duplex molecules are electrophoresed on gels, and the gels are autoradiographed. The high sensitivity and resolution of the method have made it possible to obtain a detailed map of DNase I cutting patterns in the 5' flanking sequence of the chicken adult beta (beta A)-globin gene within nuclei from various tissues. In nuclei from adult erythrocytes, this domain is hypersensitive to nucleases. However, we detect within the domain two well-defined regions that are protected against attack, indicating the presence of one or more bound factors. Nuclei from oviduct or 5-day-old embryonic erythrocytes, in which the domain is not hypersensitive, show limited and different patterns of protection.

authors

Jackson PD,Felsenfeld G

doi

10.1073/pnas.82.8.2296

subject

Has Abstract

pub_date

1985-04-01 00:00:00

pages

2296-300

issue

8

eissn

0027-8424

issn

1091-6490

journal_volume

82

pub_type

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