title: Expression level and immunolocalization of de novo methyltransferase 3 protein (TuDNMT3) in adult females and males of the two-spotted spider mite, Tetranychus urticae.
abstract: DNA methylation is an epigenetic mechanism for regulating developmental and other important processes in eukaryotes. Several essential components of the DNA methylation machinery have been identified, such as DNA methyltransferases. In the two-spotted spider mite, Tetranychus urticae Koch, we have identified one DNA methyltransferase 3 gene (Tudnmt3) and tentatively investigated its potential role in adult females and males. Here, to better elucidate the functional role of Tudnmt3, its protein structure, expression and localization were subjected to more detailed analyses. Bioinformatic analyses clearly showed that the structure of TuDNMT3 was highly conserved, with several vital amino acid residues for the activation and stabilization of its confirmation. Western blot analyses revealed that this protein was expressed in both genders, with higher expression in adult females, which was inconsistent with the gene expression, suggesting translational regulation of Tudnmt3. Subsequent immunodetection provided supportive evidence for higher expression of the TuDNMT3 protein in adult females and indicated that this protein was generally localized in the cytoplasm and that its expression was predominantly confined to the genital region of spider mites, strengthening the hypothesis that de novo methylation mediated by Tudnmt3 in gonad development or gametogenesis has a different mechanism from maintenance methyltransferase.
authors.affiliation: Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.
authors.forename: Si-Xia
authors.initials: SX
authors.lastname: Yang
authors.affiliation: Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Guo
authors.affiliation: Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.
authors.forename: Yan-Kai
authors.initials: YK
authors.lastname: Zhang
authors.affiliation: Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.
authors.forename: Jing-Tao
authors.initials: JT
authors.lastname: Sun
authors.affiliation: Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China. xyhong@njau.edu.cn.
authors.forename: Xiao-Yue
authors.initials: XY
authors.lastname: Hong
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D008297:Male; D008780:Methyltransferases / Q000235:genetics* / Q000378:metabolism*; D040102:Tetranychidae / Q000201:enzymology* / Q000235:genetics*
chemical_list: D060829:Arthropod Proteins; D008780:Methyltransferases
keywords: DNA methyltransferase 3 protein; Expression; Gender; Localization; Tetranychus urticae