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PMID: 3466161 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Nuclease resistance and the enrichment of native nuclear acceptor sites for the avian oviduct progesterone receptor.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 23 ·1986-12-00 ·Pages 8839-43

Hora J, Horton MJ, Toft DO, Spelsberg TC

Abstract

High-affinity nucleoprotein acceptor sites for the avian oviduct progesterone receptor (PR) have been enriched by a combination of nuclease digestion and centrifugation. These enriched binding elements exhibited markedly enhanced PR binding on a per mass DNA basis compared to chromatin (20- to 25-fold) or dehistonized chromatin (4- to 5-fold). Electrophoretic analysis of the nuclease-resistant DNA showed that there is a set of DNA fragments of 100-150 base pairs that are protected from digestion. Excessive digestion resulted in smaller DNA fragments and a loss of PR binding activity. The PR binding was saturable using a crude receptor preparation and displayed a competition with the same receptor preparation that was labeled with nonradioactive progesterone. The enhanced binding was also demonstrable using highly purified receptor preparations that exhibit two classes of binding sites both of which are of high affinity and saturable as assessed by Scatchard analyses. These two high-affinity classes of binding sites are shown to be competed by unlabeled purified PR. The nuclease resistance of these nucleoprotein acceptor sites from chromatin is a property similar to the nuclear matrix binding sites suggesting a relationship between these two classes of nuclear acceptor sites.

MeSH Terms
Animals Base Sequence Chickens Chromatin/ultrastructure Chromosomal Proteins, Non-Histone/metabolism DNA/metabolism DNA-Binding Proteins/metabolism Deoxyribonuclease I Deoxyribonucleoproteins/analysis Genes, Regulator Progesterone/metabolism Receptors, Progesterone/metabolism
Chemicals
Chromatin Chromosomal Proteins, Non-Histone DNA-Binding Proteins Deoxyribonucleoproteins Receptors, Progesterone Progesterone DNA Deoxyribonuclease I
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Hora J
Horton M J
Toft D O
Spelsberg T C
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41 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.83.23.8839
Published
1986-12-00
Pages
8839-43
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC387028
Subset
IM
Grants
NICHD NIH HHS · HD16705 · United States
NICHD NIH HHS · HD9140 · United States
External Links
PubMed source DOI: 10.1073/pnas.83.23.8839 PMC Full Text View journal details
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