Extracts of Drosophila Embryos Mediate Chromatin Assembly in Vitro

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1979 Nov 1

PMID 118449

Citations 20

Authors

T Nelson

T S Hsieh

D Brutlag

Affiliations

Soon will be listed here.

Abstract

Extracts of Drosophila embryos can mediate the assembly of a chromatinlike structure from histones and DNA under physiological conditions. The histone-DNA complex formed in vitro contains micrococcal nuclease-sensitive sites spaced at 200-base pair intervals. More extensive digestion of the complex by micrococcal nuclease generates 11S particles which cosediment with nucleosome core particles isolated from native chromatin. These particles contain 140-base pair DNA fragments which upon further cleavage with micrococcal nuclease give rise to a pattern of discretely sized DNA fragments characteristic of nucleosome core particles. We have assayed the chromatin assembly process both qualitatively by measuring the induction of supertwists into a relaxed circular DNA (a process requiring a nicking-closing enzyme) and quantitatively by measuring the formation of micrococcal nuclease-resistant DNA fragments from radioactively labeled linear DNA. The amount of chromatin formed depends primarily on the amount of histones, whereas the rate of assembly depends on the amount of extract protein added. The factors in the extract that mediate chromatin assembly appear to interact first with the DNA because preincubation of the DNA with the extract markedly increases the extent of assembly.

Citing Articles

Histone Gene Multiplicity and Position Effect Variegation in DROSOPHILA MELANOGASTER.

Moore G, Sinclair D, Grigliatti T Genetics. 1983; 105(2):327-44.

PMID: 17246163 PMC: 1202160. DOI: 10.1093/genetics/105.2.327.

Human histone chaperone nucleophosmin enhances acetylation-dependent chromatin transcription.

Swaminathan V, Kishore A, Febitha K, Kundu T Mol Cell Biol. 2005; 25(17):7534-45.

PMID: 16107701 PMC: 1190275. DOI: 10.1128/MCB.25.17.7534-7545.2005.

Compaction kinetics on single DNAs: purified nucleosome reconstitution systems versus crude extract.

Wagner G, Bancaud A, Quivy J, Clapier C, Almouzni G, Viovy J Biophys J. 2005; 89(5):3647-59.

PMID: 16100259 PMC: 1366857. DOI: 10.1529/biophysj.105.062786.

Linear DNA does not form chromatin containing regularly spaced nucleosomes.

Mertz J Mol Cell Biol. 1982; 2(12):1608-18.

PMID: 14582201 PMC: 369968. DOI: 10.1128/mcb.2.12.1608-1618.1982.

Human TFIIIC relieves chromatin-mediated repression of RNA polymerase III transcription and contains an intrinsic histone acetyltransferase activity.

Kundu T, Wang Z, Roeder R Mol Cell Biol. 1999; 19(2):1605-15.

PMID: 9891093 PMC: 116088. DOI: 10.1128/MCB.19.2.1605.

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