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Roles of histone H3K9 methyltransferases during Drosophila spermatogenesis.

机译:组蛋白H3K9甲基转移酶在果蝇精子发生过程中的作用。

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摘要

Epigenetic regulation of gene expression by covalent modification of histones is important for germ line cell development. In mammals, histone H3 lysine 9 (H3K9)-specific histone methyltransferases (HMTases), such as G9a, SETDB1, and SUV39H, play critical roles, but the contribution of H3K9-specific HMTases in Drosophila remains to be clarified, especially in male sperm. Here, we performed immunocytochemical analyses with a specific antibody to dG9a, Drosophila G9a ortholog, and demonstrated localization in the cytoplasm from the growth to elongation stages of spermatogenesis. In the subsequent early canoe stage, strong dG9a signals were detected exclusively in nuclei, suggesting a regulatory role. However, mono-, di-, and trimethylated H3K9 signals were not extensively decreased in a homozygous dG9a null mutant throughout these stages. In contrast, mono- and trimethylated H3K9 signals were extensively decreased in a heterozygous DmSetdb1 mutant during spermatogenesis, and similar reduction in monomethylated H3K9 signals was observed in a homozygous Su(var)3-9 mutant. Therefore, DmSETDB1 is likely to be mainly responsible for mono- and trimethylation of H3K9 and SU(VAR)3-9 for monomethylation of H3K9 during spermatogenesis. However, the reduced methylation of H3K9 in premeiotic spermatocytes did not influence X-Y chromosome disjunction in male meiosis, suggesting that it may not be critical for spermatogenesis in Drosophila.
机译:通过组蛋白的共价修饰对基因表达进行表观遗传调控对于种系细胞发育很重要。在哺乳动物中,组蛋白H3赖氨酸9(H3K9)特异的组蛋白甲基转移酶(HMTase),例如G9a,SETDB1和SUV39H,起着关键作用,但H3K9特异HMTases在果蝇中的作用仍有待阐明,尤其是在男性精子中。在这里,我们用针对dG9a,果蝇G9a直系同源物的特异性抗体进行了免疫细胞化学分析,并证明了从生精到生精期的细胞质中的定位。在随后的独木舟早期,仅在细胞核中检测到强dG9a信号,表明其具有调节作用。但是,在这些阶段的纯合dG9a无效突变体中,单甲基化,二甲基化和三甲基化的H3K9信号并未广泛降低。相反,在精子发生过程中,杂合的DmSetdb1突变体中的单甲基化和三甲基化的H3K9信号被大量降低,而纯合的Su(var)3-9突变体中的单甲基化的H3K9信号也被类似地降低。因此,DmSETDB1可能主要负责H3K9的单甲基化和三甲基化,而SU(VAR)3-9则负责精子发生过程中H3K9的单甲基化。但是,减数分裂前的精母细胞中H3K9甲基化的减少不会影响雄性减数分裂中X-Y染色体的分离,这表明它对果蝇的精子发生不是至关重要的。

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