A comparison of DNA repair using the comet assay in tobacco seedlings after exposure to alkylating agents or ionizing radiation

Tomáš Gichner, Ondřej Ptáček, Diana A. Stavreva, Elizabeth D. Wagner, Michael Jacob Plewa

Research output: Contribution to journalArticle

Abstract

We employed single cell gel electrophoresis to analyze the kinetics of DNA repair in nuclei isolated from tobacco plants exposed to ethyl methanesulfonate (EMS), N-ethyl-N-nitrosourea (ENU) and γ-radiation. DNA repair was measured as the reduction of the tail moment values as a function of time after the mutagen treatment ended. DNA damage in leaf nuclei of EMS- or ENU-treated tobacco plants persisted over a 72 h recovery period. However, a reduction of the SCGE tail moment values in nuclei isolated from leaves was observed over a 4-week period of recovery. Newly emerged leaves expressed a lower level of DNA damage due to more efficient repair and/or dilution of initial DNA lesions during cell division. After 24 h recovery, leaf nuclei from cells exposed to 20 or 40 Gy of γ-radiation expressed complete DNA repair. These data indicate that DNA lesions induced by alkylating agents are not readily repaired and persist beyond 4 weeks. Enzymes necessary to repair γ-induced DNA lesions are fully functional in non-replicating leaf cells and single and double strand breaks are rapidly repaired. (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)1-9
Number of pages9
JournalMutation Research - Genetic Toxicology and Environmental Mutagenesis
Volume470
Issue number1
DOIs
StatePublished - Oct 10 2000

Fingerprint

Comet Assay
Alkylating Agents
Ionizing Radiation
Seedlings
DNA Repair
Tobacco
Ethyl Methanesulfonate
Ethylnitrosourea
DNA Damage
Radiation Dosage
DNA
Mutagens
Cell Nucleus
Cell Division
Radiation
Enzymes

Keywords

  • Comet assay
  • EMS
  • ENU
  • Ethyl methanesulfonate
  • N-ethyl-N-nitrosourea
  • Nicotiana tabacum var. xanthi
  • SCGE
  • Single cell gel electrophoresis
  • γ-radiation

ASJC Scopus subject areas

  • Genetics
  • Health, Toxicology and Mutagenesis

Cite this

A comparison of DNA repair using the comet assay in tobacco seedlings after exposure to alkylating agents or ionizing radiation. / Gichner, Tomáš; Ptáček, Ondřej; Stavreva, Diana A.; Wagner, Elizabeth D.; Plewa, Michael Jacob.

In: Mutation Research - Genetic Toxicology and Environmental Mutagenesis, Vol. 470, No. 1, 10.10.2000, p. 1-9.

Research output: Contribution to journalArticle

Gichner, Tomáš ; Ptáček, Ondřej ; Stavreva, Diana A. ; Wagner, Elizabeth D. ; Plewa, Michael Jacob. / A comparison of DNA repair using the comet assay in tobacco seedlings after exposure to alkylating agents or ionizing radiation. In: Mutation Research - Genetic Toxicology and Environmental Mutagenesis. 2000 ; Vol. 470, No. 1. pp. 1-9.
@article{76ea42b169894c1080197f384cdbac9b,
title = "A comparison of DNA repair using the comet assay in tobacco seedlings after exposure to alkylating agents or ionizing radiation",
abstract = "We employed single cell gel electrophoresis to analyze the kinetics of DNA repair in nuclei isolated from tobacco plants exposed to ethyl methanesulfonate (EMS), N-ethyl-N-nitrosourea (ENU) and γ-radiation. DNA repair was measured as the reduction of the tail moment values as a function of time after the mutagen treatment ended. DNA damage in leaf nuclei of EMS- or ENU-treated tobacco plants persisted over a 72 h recovery period. However, a reduction of the SCGE tail moment values in nuclei isolated from leaves was observed over a 4-week period of recovery. Newly emerged leaves expressed a lower level of DNA damage due to more efficient repair and/or dilution of initial DNA lesions during cell division. After 24 h recovery, leaf nuclei from cells exposed to 20 or 40 Gy of γ-radiation expressed complete DNA repair. These data indicate that DNA lesions induced by alkylating agents are not readily repaired and persist beyond 4 weeks. Enzymes necessary to repair γ-induced DNA lesions are fully functional in non-replicating leaf cells and single and double strand breaks are rapidly repaired. (C) 2000 Elsevier Science B.V.",
keywords = "Comet assay, EMS, ENU, Ethyl methanesulfonate, N-ethyl-N-nitrosourea, Nicotiana tabacum var. xanthi, SCGE, Single cell gel electrophoresis, γ-radiation",
author = "Tom{\'a}š Gichner and Ondřej Pt{\'a}ček and Stavreva, {Diana A.} and Wagner, {Elizabeth D.} and Plewa, {Michael Jacob}",
year = "2000",
month = "10",
day = "10",
doi = "10.1016/S1383-5718(00)00081-4",
language = "English (US)",
volume = "470",
pages = "1--9",
journal = "Mutation Research - Genetic Toxicology and Environmental Mutagenesis",
issn = "1383-5718",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - A comparison of DNA repair using the comet assay in tobacco seedlings after exposure to alkylating agents or ionizing radiation

AU - Gichner, Tomáš

AU - Ptáček, Ondřej

AU - Stavreva, Diana A.

AU - Wagner, Elizabeth D.

AU - Plewa, Michael Jacob

PY - 2000/10/10

Y1 - 2000/10/10

N2 - We employed single cell gel electrophoresis to analyze the kinetics of DNA repair in nuclei isolated from tobacco plants exposed to ethyl methanesulfonate (EMS), N-ethyl-N-nitrosourea (ENU) and γ-radiation. DNA repair was measured as the reduction of the tail moment values as a function of time after the mutagen treatment ended. DNA damage in leaf nuclei of EMS- or ENU-treated tobacco plants persisted over a 72 h recovery period. However, a reduction of the SCGE tail moment values in nuclei isolated from leaves was observed over a 4-week period of recovery. Newly emerged leaves expressed a lower level of DNA damage due to more efficient repair and/or dilution of initial DNA lesions during cell division. After 24 h recovery, leaf nuclei from cells exposed to 20 or 40 Gy of γ-radiation expressed complete DNA repair. These data indicate that DNA lesions induced by alkylating agents are not readily repaired and persist beyond 4 weeks. Enzymes necessary to repair γ-induced DNA lesions are fully functional in non-replicating leaf cells and single and double strand breaks are rapidly repaired. (C) 2000 Elsevier Science B.V.

AB - We employed single cell gel electrophoresis to analyze the kinetics of DNA repair in nuclei isolated from tobacco plants exposed to ethyl methanesulfonate (EMS), N-ethyl-N-nitrosourea (ENU) and γ-radiation. DNA repair was measured as the reduction of the tail moment values as a function of time after the mutagen treatment ended. DNA damage in leaf nuclei of EMS- or ENU-treated tobacco plants persisted over a 72 h recovery period. However, a reduction of the SCGE tail moment values in nuclei isolated from leaves was observed over a 4-week period of recovery. Newly emerged leaves expressed a lower level of DNA damage due to more efficient repair and/or dilution of initial DNA lesions during cell division. After 24 h recovery, leaf nuclei from cells exposed to 20 or 40 Gy of γ-radiation expressed complete DNA repair. These data indicate that DNA lesions induced by alkylating agents are not readily repaired and persist beyond 4 weeks. Enzymes necessary to repair γ-induced DNA lesions are fully functional in non-replicating leaf cells and single and double strand breaks are rapidly repaired. (C) 2000 Elsevier Science B.V.

KW - Comet assay

KW - EMS

KW - ENU

KW - Ethyl methanesulfonate

KW - N-ethyl-N-nitrosourea

KW - Nicotiana tabacum var. xanthi

KW - SCGE

KW - Single cell gel electrophoresis

KW - γ-radiation

UR - http://www.scopus.com/inward/record.url?scp=0343714603&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0343714603&partnerID=8YFLogxK

U2 - 10.1016/S1383-5718(00)00081-4

DO - 10.1016/S1383-5718(00)00081-4

M3 - Article

C2 - 10986470

AN - SCOPUS:0343714603

VL - 470

SP - 1

EP - 9

JO - Mutation Research - Genetic Toxicology and Environmental Mutagenesis

JF - Mutation Research - Genetic Toxicology and Environmental Mutagenesis

SN - 1383-5718

IS - 1

ER -