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Titolo:
STUDIES ON THE ISOLATED TRANSCRIPTIONALLY ACTIVE AND INACTIVE CHROMATIN FRACTIONS FROM RAT-LIVER NUCLEI
Autore:
YU FL; BENDER W;
Indirizzi:
UNIV ILLINOIS,COLL MED,DEPT BIOMED SCI,1601 PKVIEW AVE ROCKFORD IL 61107
Titolo Testata:
Journal of biochemical and biophysical methods
fascicolo: 1, volume: 30, anno: 1995,
pagine: 21 - 36
SICI:
0165-022X(1995)30:1<21:SOTITA>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
DEPENDENT RNA-SYNTHESIS; AFLATOXIN-B1 INHIBITION; PREFERENTIAL BINDING; NUCLEOSOME STRUCTURE; CHICKEN ERYTHROCYTE; SYNTHESIS INVITRO; HEPATIC NUCLEAR; GENE-SEQUENCES; HISTONE H-1; DNA;
Keywords:
ACTIVE CHROMATIN; HISTONE/HISTONE-LIKE PROTEIN; NUCLEAR/NUCLEOLAR RNA SYNTHESIS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
74
Recensione:
Indirizzi per estratti:
Citazione:
F.L. Yu e W. Bender, "STUDIES ON THE ISOLATED TRANSCRIPTIONALLY ACTIVE AND INACTIVE CHROMATIN FRACTIONS FROM RAT-LIVER NUCLEI", Journal of biochemical and biophysical methods, 30(1), 1995, pp. 21-36

Abstract

Using mild sonication, nucleoplasmic, nucleolar, and subnucleolar P-3and S-3 chromatin fractions are isolated from rat liver nuclei. Thesefractions differ widely (over 80-fold) from each other in transcriptional activity as measured by the chromatin bound engaged RNA polymerases. Chemical analyses indicate that the active chromatin, e.g. P-3 andnucleolar fractions, are rich in RNA and protein as compared to the inactive chromatin, e.g. nucleoplasmic, and S-3 fractions. However, theDNA base content are all the same, showing 40% GC and 60% AT, including P-3 which is enriched in rDNA. Polyacrylamide gel electrophoresis of the 0.25 N HCl extracted proteins shows that all five histones are present in active chromatin. Additionally, the gel reveals two protein bands, one ahead of histone H2B and another ahead of histone H4, that are diminished or missing from the inactive chromatin. On the other hand, there is a fast moving protein band ahead of H4 in the inactive chromatin that is almost absent in the active chromatin. Transcriptionaltests using E. coli RNA polymerase and several synthetic DNA templates of known base content and sequence indicate that the 0.25 N HCl soluble protein extracts from active chromatin contain activator proteins which are capable of countering the histone suppressors present in theextracts in a DNA base and sequence specific manner. The data show that although the histone suppressors are able to strongly inhibit the template function of poly[d(A-T)], the protein activators are able to overcome the suppressor activity and stimulate RNA synthesis several-fold when poly(dA).poly(dT) or poly(dT) is used.

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Documento generato il 30/11/20 alle ore 07:23:11