title: ddm1 plants are sensitive to methyl methane sulfonate and NaCl stresses and are deficient in DNA repair.
abstract: Plant response to stress includes changes in gene expression and chromatin structure. Our previous work showed that Arabidopsis thaliana Dicer-like (DCL) mutants were impaired in transgenerational response to stress that included an increase in recombination frequency, cytosine methylation and stress tolerance. It can be hypothesized that changes in chromatin structure are important for an efficient stress response. To test this hypothesis, we analyzed the stress response of ddm1, a mutant impaired in DDM1, a member of the SWI/SNF family of adenosine triphosphate-dependent chromatin remodeling genes. We exposed Arabidopsis thaliana ddm1 mutants to methyl methane sulfonate (MMS) and NaCl and found that these plants were more sensitive. At the same time, ddm1 plants were similar to wild-type plants in sensitivity to temperature and bleomycin stresses. Direct comparison to met1 plants, deficient in maintenance methyltransferase MET1, showed higher sensitivity of ddm1 plants to NaCl. The level of DNA strand breaks upon exposure to MMS increased in wild-type plants but decreased in ddm1 plants. DNA methylation analysis showed that heterozygous ddm1/DDM1 plants had lower methylation as compared to fourth generation of homozygous ddm1/ddm1 plants. Exposure to MMS resulted in a decrease in methylation in wild-type plants and an increase in ddm1 plants. Finally, in vitro DNA excision repair assay showed lower capacity for ddm1 mutant. Our results provided a new example of a link between genetic genome stability and epigenetic genome stability.
  
  
  CONCLUSIONS
  We demonstrate that heterozygous ddm1/DDM1 plants are more sensitive to stress and have more severe changes in methylation than homozygous ddm1/ddm1 plants.
authors.affiliation: Department of Biological Sciences, University of Lethbridge, University Drive 4401, Lethbridge, AB, T1K 3M4, Canada. youli.yao@uleth.ca
authors.forename: Youli
authors.initials: Y
authors.lastname: Yao
authors.forename: Andriy
authors.initials: A
authors.lastname: Bilichak
authors.forename: Andrey
authors.initials: A
authors.lastname: Golubov
authors.forename: Igor
authors.initials: I
authors.lastname: Kovalchuk
mesh_terms: D000222:Adaptation, Physiological / Q000187:drug effects; D017360:Arabidopsis / Q000187:drug effects* / Q000502:physiology*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D001681:Biological Assay; D001761:Bleomycin / Q000494:pharmacology; D003433:Crosses, Genetic; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects; D019175:DNA Methylation / Q000187:drug effects / Q000235:genetics; D004260:DNA Repair / Q000187:drug effects*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006579:Heterozygote; D008741:Methyl Methanesulfonate / Q000494:pharmacology*; D009154:Mutation / Q000235:genetics; D015203:Reproducibility of Results; D012965:Sodium Chloride / Q000494:pharmacology*; D013312:Stress, Physiological / Q000187:drug effects*; D013696:Temperature; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; C118957:DDM1 protein, Arabidopsis; D004268:DNA-Binding Proteins; D014157:Transcription Factors; D001761:Bleomycin; D012965:Sodium Chloride; D008741:Methyl Methanesulfonate; C499608:MET1 protein, Arabidopsis; D004248:DNA (Cytosine-5-)-Methyltransferases