The nuclease sensitivity of active genes

Brief mlcrococcal nuclease digestion of chick embryonic red blood cells results in preferential excision and solubilization of monomer nucleosomes associated with g-globin sequences and also 5’-sequences flanking the g-globin gene. Both regions are DNAse-I sensitive in nuclei. Such salt-soluble nucl...

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Bibliographic Details
Published in:Nucleic acids research 1983-02, Vol.11 (3), p.753-772
Main Authors: Nicolas, Robert H., Wright, Carol A., Cockerill, Peter N., Wyke, John A., Goodwin, Graham H.
Format: Article
Language:English
Subjects:
Animals
avian sarcoma virus
Avian Sarcoma Viruses - genetics
Base Sequence
Cell Transformation, Neoplastic
Chickens
Deoxyribonuclease I
DNA - blood
Endodeoxyribonucleases
Erythrocytes - metabolism
Genes
Globins - genetics
Micrococcal Nuclease - metabolism
Nucleic Acid Hybridization
Nucleosomes - metabolism
Ovalbumin - genetics
Rats
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Summary:Brief mlcrococcal nuclease digestion of chick embryonic red blood cells results in preferential excision and solubilization of monomer nucleosomes associated with g-globin sequences and also 5’-sequences flanking the g-globin gene. Both regions are DNAse-I sensitive in nuclei. Such salt-soluble nucleosomes are enriched in all four major HMG proteins but HMG1 and 2 are only weakly associated. These nucleosomes appear to have lost much of the DNAse-I sensitivity of active genes. The HMG14 and 17-containing salt-soluble nucleosomes separated by electrophoresis are not DNAse-I sensitive and contain inactive gene sequences as well as active sequences. Reconstitution of HMG proteins onto bulk nucleosomes or chromatin failed to reveal an HMG-dependent sensitivity of active genes as assayed by dot-blot hybridization and it was found that the DNAse-I sensitivity of ASV proviral sequences as assayed by dot-blot hybridization was not HMG-dependent. These results indicate that higher order chromatin structures might be responsible for nuclease sensitivity of active genes.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/11.3.753