Philipps-Universität Marburg, Fachbereich Biologie, Entwicklungsbiologie , Karl-von-Frisch Strasse 8, 35043 Marburg , Germany.
Biol Open. 2012 Jun 15;1(6):597-606. doi: 10.1242/bio.20121255. Epub 2012 May 9.
By a conserved cellular differentiation process, spermatogenesis leads to formation of haploid sperm for successful reproduction. In Drosophila and in mammals, post-meiotic spermatid differentiation depends on several translationally repressed and stored mRNAs that are often expressed exclusively in the testis through a cell type specific transcriptional program. In Drosophila, the mRNAs of proteins required for post-meiotic chromatin reorganisation, like ProtB and Mst77F, are transcribed in meiotic spermatocytes and subjected to translational repression for days. Transcription of many of these translationally repressed mRNAs depends on testis-specific homologs of TATA box binding protein-associated factors (tTAFs). Here, we identified the testis-specific bromodomain protein, tBRD-1, that is only expressed in primary spermatocytes. Bromodomain proteins are able to recognise and bind acetylated histones and non-histone proteins. We generated tbrd-1 mutant flies and observed that function of tBRD-1 is required for male fertility. tBRD-1 partially colocalised with tTAFs, TAF1 and Polycomb to a Fibrillarin-deficient region within the spermatocyte nucleolus. The nucleolar localisation of tBRD-1 depended on tTAF function but not the other way round. Further, we could show that ectopically expressed tBRD-1-eGFP is able to bind to the interbands of polytene chromosomes. By inhibitor treatment of cultured testis we observed that sub-cellular localisation of tBRD-1 may depend on the acetylation status of primary spermatocytes.
通过保守的细胞分化过程,精子发生导致形成单倍体精子以实现成功繁殖。在果蝇和哺乳动物中,减数后精母细胞的分化依赖于几种翻译后被抑制和储存的 mRNA,这些 mRNA 通常通过特定于细胞类型的转录程序在睾丸中特异性表达。在果蝇中,后减数期染色质重组所需的蛋白质的 mRNAs,如 ProtB 和 Mst77F,在减数分裂精母细胞中转录,并被翻译抑制数天。许多这些翻译后被抑制的 mRNAs 的转录依赖于睾丸特异性 TATA 盒结合蛋白相关因子(tTAFs)的同源物。在这里,我们鉴定了仅在初级精母细胞中表达的睾丸特异性溴结构域蛋白 tBRD-1。溴结构域蛋白能够识别和结合乙酰化的组蛋白和非组蛋白。我们生成了 tbrd-1 突变体果蝇,并观察到 tBRD-1 的功能对于雄性生育力是必需的。tBRD-1 与 tTAFs、TAF1 和 Polycomb 部分共定位到精母细胞核仁中 Fibrillarin 缺陷区域。tBRD-1 的核仁定位依赖于 tTAF 功能,但反之则不然。此外,我们可以证明异位表达的 tBRD-1-eGFP 能够结合多线染色体的间带。通过培养的睾丸的抑制剂处理,我们观察到 tBRD-1 的亚细胞定位可能取决于初级精母细胞的乙酰化状态。