title: Changes in DNA methylation and transgenerational mobilization of a transposable element (mPing) by the topoisomerase II inhibitor, etoposide, in rice.
abstract: BACKGROUND
  Etoposide (epipodophyllotoxin) is a chemical commonly used as an anti-cancer drug which inhibits DNA synthesis by blocking topoisomerase II activity. Previous studies in animal cells have demonstrated that etoposide constitutes a genotoxic stress which may induce genomic instability including mobilization of normally quiescent transposable elements (TEs). However, it remained unknown whether similar genetically mutagenic effects could be imposed by etoposide in plant cells. Also, no information is available with regard to whether the drug may cause a perturbation of epigenetic stability in any organism.
  
  
  RESULTS
  To investigate whether etoposide could generate genetic and/or epigenetic instability in plant cells, we applied etoposide to germinating seeds of six cultivated rice (Oryza sativa L.) genotypes including both subspecies, japonica and indica. Based on the methylation-sensitive gel-blotting results, epigenetic changes in DNA methylation of three TEs (Tos17, Osr23 and Osr36) and two protein-encoding genes (Homeobox and CDPK-related genes) were detected in the etoposide-treated plants (S0 generation) in four of the six studied japonica cultivars, Nipponbare, RZ1, RZ2, and RZ35, but not in the rest japonica cultivar (Matsumae) and the indica cultivar (93-11). DNA methylation changes in the etoposide-treated S0 rice plants were validated by bisulfite sequencing at both of two analyzed loci (Tos17 and Osr36). Transpositional activity was tested for eight TEs endogenous to the rice genome in both the S0 plants and their selfed progenies (S1 and S2) of one of the cultivars, RZ1, which manifested heritable phenotypic variations. Results indicated that no transposition occurred in the etoposide-treated S0 plants for any of the TEs. Nonetheless, a MITE transposon, mPing, showed rampant mobilization in the S1 and S2 progenies descended from the drug-treated S0 plants.
  
  
  CONCLUSIONS
  Our results demonstrate that etoposide imposes a similar genotoxic stress on plant cells as it does on animal and human cells, which may induce transgenerational genomic instability by instigating transpositional activation of otherwise dormant TEs. In addition, we show for the first time that etoposide may induce epigenetic instability in the form of altered DNA methylation patterns in eukaryotes. However, penetrance of the genotoxic effects of etoposide on plant cells, as being reflected as genetic and epigenetic instability, appears to be in a strictly genotype- and/or generation-dependent manner.
authors.affiliation: Key Laboratory of Molecular Epigenetics of MOE, and Institute of Genetics & Cytology, Northeast Normal University, Changchun 130024, China.
authors.forename: Xuejiao
authors.initials: X
authors.lastname: Yang
authors.forename: Yingjie
authors.initials: Y
authors.lastname: Yu
authors.forename: Lily
authors.initials: L
authors.lastname: Jiang
authors.forename: Xiuyun
authors.initials: X
authors.lastname: Lin
authors.forename: Chunyu
authors.initials: C
authors.lastname: Zhang
authors.forename: Xiufang
authors.initials: X
authors.lastname: Ou
authors.forename: Kenji
authors.initials: K
authors.lastname: Osabe
authors.forename: Bao
authors.initials: B
authors.lastname: Liu
mesh_terms: D001483:Base Sequence; D003596:Cytosine / Q000378:metabolism; D019175:DNA Methylation; D004251:DNA Transposable Elements; D044127:Epigenesis, Genetic; D005047:Etoposide / Q000494:pharmacology*; D017343:Genes, Plant; D056426:Genetic Loci; D042822:Genomic Instability; D005838:Genotype; D040582:Inheritance Patterns; D012275:Oryza / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D019683:Penetrance; D059828:Plant Cells / Q000187:drug effects / Q000378:metabolism; D012639:Seeds / Q000187:drug effects / Q000235:genetics; D057945:Self-Fertilization; D059005:Topoisomerase II Inhibitors / Q000494:pharmacology; D015533:Transcriptional Activation
chemical_list: D004251:DNA Transposable Elements; D059005:Topoisomerase II Inhibitors; D005047:Etoposide; D003596:Cytosine