The Influence of Chromatin Structure on the Distribution of DNA Repair Synthesis Studied by Nuclease Digestion

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1979 Jan 1

PMID 424297

Citations 6

Authors

W J Bodell

M R BANERJEE

Affiliations

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Abstract

The influence of chromatin structure on the distribution of DNA repair synthesis was studied by enzymatic digestion of "repair labeled" nuclei of mouse mammary cells: "repair labeled" nuclei were isolated from pregnancy mammary tissue fragments, treated in vitro with methylmethanesulfonate (MMS) or methylnitrosourea (MNU), and pulse-labeled with 3H-thymidine in the presence of hydroxyurea in the culture medium. Micrococcal nuclease digestion of "repair labeled" nuclei indicates that at early hours after treatment with the alkylating agents 70-80% of the total repair synthesis is located in the linker portion of the nucleosome. However, 6-12 hours after treatment DNA repair synthesis is more evenly distributed throughout the core and linker portion of the nucleosome. "Repair labeled" mammary cell nuclei were also digested with DNase I under conditions selective for transcriptionally active chromatin. A two-fold higher level of repair synthesis was found in the transcriptionally active chromatin of "repair labeled" nuclei isolated from MMS or MNU treated mammary fragments, pulse-labeled at different times after treatment. The results indicate that structural constitution of the chromatin may influence the distribution of DNA repair synthesis both at the nucleosome level, and at higher levels of chromatin organization. This may be due to 1) nonrandom base alkylation in chromatin or 2) areas in chromatin with increased accessibility for the repair enzymes to the alkylated bases.

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The use of radioimmunoassay to study the formation and disappearance of O6-methylguanine in mouse liver satellite and main-band DNA following dimethylnitrosamine administration.

Kyrtopoulos S, Swann P J Cancer Res Clin Oncol. 1980; 98(2):127-38.

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Transient conformation changes in chromatin during excision repair of ultraviolet damage to DNA.

Bodell W, Cleaver J Nucleic Acids Res. 1981; 9(1):203-13.

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Non-random effect on RNA synthesis in liver chromatin by administration of dimethylnitrosamine to mice.

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Distribution of DNA damage in chromatin and its relation to repair in human cells treated with 7-bromomethylbenz(a) anthracene.

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