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PMID: 6743285 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A chromosomal phosphoprotein is preferentially released by mild micrococcal-nuclease digestion.

The Biochemical journal ·Vol. 220 ·No. 2 ·1984-06-01 ·Pages 539-45

Liew CC, Halikowski MJ, Zhao MS

Abstract

[32P]Pi was administered to rats (5mCi/rat) 2h before the isolation of liver nuclei. The isolated nuclei were subjected to mild micrococcal-nuclease digestion for 2.5, 5 and 10 min at 37 degrees C, and the mononucleosomal fraction was subsequently isolated by sucrose-density-gradient centrifugation. The specific radioactivity of 32P-labelled mononucleosomal fractions decreased with increased digestion times. A phosphorylated chromosomal protein, B2 (Mr 68000, pI6.5-8.2), was demonstrated immunologically in the mononucleosomal fraction by using an antibody specific to this electrophoretically purified phosphoprotein. The incorporation of 32P into this phosphoprotein, previously shown to be mainly through covalent linkage, was revealed by antibody precipitation followed by gel electrophoresis. The rate of release of acid-soluble nucleotides by micrococcal-nuclease digestion of liver nuclei from partially hepatectomized rats 16 h after operation was strikingly higher than that for sham-operated controls. After partial hepatectomy, an increase in 32P incorporation into phosphoprotein in the monomer fractions specifically precipitated by this antibody was also found. This suggests that the phosphorylated non-histone chromatin protein B2 is preferentially associated with the transcriptionally active chromatin.

MeSH Terms
Animals Centrifugation, Density Gradient Chemical Precipitation Chromosomal Proteins, Non-Histone/immunology,metabolism Electrophoresis, Polyacrylamide Gel Hepatectomy In Vitro Techniques Male Micrococcal Nuclease/pharmacology Nucleosomes/drug effects,metabolism Nucleotides Phosphates/metabolism Phosphoproteins/immunology,metabolism Rats Rats, Inbred Strains
Chemicals
Chromosomal Proteins, Non-Histone Nucleosomes Nucleotides Phosphates Phosphoproteins nuclear phosphoprotein B2, rat Micrococcal Nuclease
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Liew C C
Halikowski M J
Zhao M S
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2200539
Published
1984-06-01
Pages
539-45
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1153657
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2200539 PMC Full Text View journal details
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