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Removal of pyrimidine dimers from Saccharomyces cerevisiae nuclear DNA under nongrowth conditions as detected by a sensitive, enzymatic assay

Journal Article:

Abstract

A sensitive and quantitative procedure for the detection of pyrimidine dimers in yeast nuclear DNA is described. The assay employs dimer-specific, endonuclease activities from Micrococcus luteus together with DNA sedimentation through calibrated, alkaline sucrose gradients to detect endonuclease-induced, single-strand breaks. Breaks were induced in a dose-dependent manner from 0 to 80 J m/sup -2/ at 254 nm and in numbers equivalent to the numbers of dimers induced by similar doses. Endonuclease-sensitive sites in the wild-type, haploid strain S288C, after irradiation with 5 J m/sup -2/ (254 nm), were removed in less than 5 min when cells were incuba ted in buffer (pH 7.0) at 28/sup 0/C. After irra diation with dos es from 30 to 100 J m/sup -2/ site removal in S288C required longer postirradiation incubations and was about 90% complete. In a radiation-sensitive strain carrying the mutant allele rad 4-3 the number of endonuclease-sensitive sites remained constant for 6 h after irradiation with 5 J m/sup -2/. The retention of sites in this strain indicates that it is defective in the excision of pyrimidine dimers. (Auth.
Authors:
Reynolds, R J [1] 
  1. Tennessee Univ., Oak Ridge (USA). Graduate School of Biomedical Sciences
Publication Date:
Apr 01, 1978
Product Type:
Journal Article
Reference Number:
AIX-09-395034; EDB-78-110800
Resource Relation:
Journal Name: Mutat. Res.; (Netherlands); Journal Volume: 50:1
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; DNA; BIOLOGICAL REPAIR; PYRIMIDINES; BIOASSAY; BIOLOGICAL LOCALIZATION; DIMERS; DOSE-RESPONSE RELATIONSHIPS; FAR ULTRAVIOLET RADIATION; LOW DOSE IRRADIATION; NUCLEASES; SACCHAROMYCES CEREVISIAE; SEDIMENTATION; AZINES; BIOLOGICAL RECOVERY; BIOMASS; ELECTROMAGNETIC RADIATION; ENERGY SOURCES; ENZYMES; FUNGI; HETEROCYCLIC COMPOUNDS; IRRADIATION; MICROORGANISMS; NUCLEIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PHOSPHOTRANSFERASES; PLANTS; RADIATIONS; RECOVERY; RENEWABLE ENERGY SOURCES; REPAIR; SACCHAROMYCES; TRANSFERASES; ULTRAVIOLET RADIATION; YEASTS; 560112* - Radiation Effects on Biochemicals- In Microorganisms- (-1987)
OSTI ID:
7087405
Country of Origin:
Netherlands
Language:
English
Other Identifying Numbers:
Journal ID: CODEN: MUREA
Submitting Site:
INIS
Size:
Pages: 43-56
Announcement Date:

Journal Article:

Citation Formats

Reynolds, R J. Removal of pyrimidine dimers from Saccharomyces cerevisiae nuclear DNA under nongrowth conditions as detected by a sensitive, enzymatic assay. Netherlands: N. p., 1978. Web.
Reynolds, R J. Removal of pyrimidine dimers from Saccharomyces cerevisiae nuclear DNA under nongrowth conditions as detected by a sensitive, enzymatic assay. Netherlands.
Reynolds, R J. 1978. "Removal of pyrimidine dimers from Saccharomyces cerevisiae nuclear DNA under nongrowth conditions as detected by a sensitive, enzymatic assay." Netherlands.
@misc{etde_7087405,
title = {Removal of pyrimidine dimers from Saccharomyces cerevisiae nuclear DNA under nongrowth conditions as detected by a sensitive, enzymatic assay}
author = {Reynolds, R J}
abstractNote = {A sensitive and quantitative procedure for the detection of pyrimidine dimers in yeast nuclear DNA is described. The assay employs dimer-specific, endonuclease activities from Micrococcus luteus together with DNA sedimentation through calibrated, alkaline sucrose gradients to detect endonuclease-induced, single-strand breaks. Breaks were induced in a dose-dependent manner from 0 to 80 J m/sup -2/ at 254 nm and in numbers equivalent to the numbers of dimers induced by similar doses. Endonuclease-sensitive sites in the wild-type, haploid strain S288C, after irradiation with 5 J m/sup -2/ (254 nm), were removed in less than 5 min when cells were incuba ted in buffer (pH 7.0) at 28/sup 0/C. After irra diation with dos es from 30 to 100 J m/sup -2/ site removal in S288C required longer postirradiation incubations and was about 90% complete. In a radiation-sensitive strain carrying the mutant allele rad 4-3 the number of endonuclease-sensitive sites remained constant for 6 h after irradiation with 5 J m/sup -2/. The retention of sites in this strain indicates that it is defective in the excision of pyrimidine dimers. (Auth.}
journal = {Mutat. Res.; (Netherlands)}
volume = {50:1}
journal type = {AC}
place = {Netherlands}
year = {1978}
month = {Apr}
}