Metcalf Chad E, Wassarman David A
University of Wisconsin School of Medicine and Public Health, Department of Biomolecular Chemistry, Madison, Wisconsin 53706, USA.
Dev Dyn. 2007 Oct;236(10):2836-43. doi: 10.1002/dvdy.21294.
In Drosophila, testis-specific TBP-associated factors (tTAFs) predominantly localize to spermatocyte nucleoli and regulate the transcription of genes necessary for spermatocyte entry into meiosis. tTAFs are paralogs of generally expressed TAF subunits of transcription factor IID (TFIID). Our recent observation that the generally expressed TAF1 isoform TAF1-2 is greatly enriched in testes prompted us to explore the functional relationship between general TAFs and tTAFs during spermatogenesis. Analysis by immunofluorescence microscopy revealed that among the general TFIID subunits examined (TATA-box binding protein [TBP], TAF1, TAF4, TAF5, and TAF9), only TAF1 colocalized with the tTAF Mia in spermatocyte nucleoli. Nucleolar localization of TAF1, but not Mia, was disrupted in tTAF mutant flies, and TAF1 dissociated from DNA prior to Mia as spermatocytes entered meiosis. Taken together, our results suggest stepwise assembly of a testis-specific TFIID complex (tTFIID) whereby a TAF1 isoform, presumably TAF1-2, is recruited to a core subassembly of tTAFs in spermatocyte nucleoli.
在果蝇中,睾丸特异性TBP相关因子(tTAFs)主要定位于精母细胞的核仁,并调节精母细胞进入减数分裂所需基因的转录。tTAFs是转录因子IID(TFIID)中一般表达的TAF亚基的旁系同源物。我们最近观察到一般表达的TAF1异构体TAF1-2在睾丸中高度富集,这促使我们探索在精子发生过程中一般TAFs与tTAFs之间的功能关系。免疫荧光显微镜分析显示,在所检测的一般TFIID亚基(TATA盒结合蛋白[TBP]、TAF1、TAF4、TAF5和TAF9)中,只有TAF1与精母细胞核仁中的tTAF Mia共定位。在tTAF突变果蝇中,TAF1的核仁定位被破坏,但Mia不受影响,并且随着精母细胞进入减数分裂,TAF1在Mia之前从DNA上解离。综上所述,我们的结果表明睾丸特异性TFIID复合物(tTFIID)是逐步组装的,其中一种TAF1异构体,可能是TAF1-2,被招募到精母细胞核仁中tTAFs的核心亚组件上。