Richler C, Ast G, Goitein R, Wahrman J, Sperling R, Sperling J
Department of Genetics, Hebrew University of Jerusalem, Israel.
Mol Biol Cell. 1994 Dec;5(12):1341-52. doi: 10.1091/mbc.5.12.1341.
The study of the effect of programmed cessation of transcription in a large nuclear domain, on the distribution of elements of the pre-mRNA splicing machinery, is the main aim of this paper. To this end, we took advantage of the nuclear partitioning of mouse spermatocytes early in meiosis into autosomal transcribing and XY nontranscribing compartments. This system also allows to extend this study to stages in sperm differentiation that are accompanied by reduction and eventual cessation of transcription. We show by indirect immunofluorescence in spermatogenetic cells that 1) fluorescent signals of the pre-mRNA splicing factors SF53/4 and SC35, of the Sm antigens, and of RNA polymerase II, are largely absent from the nontranscribing, X-inactivated compartment, but are abundantly present in the transcribing autosomal compartment and 2) the presence, gradual reduction, and absence of transcriptive activity in nuclei undergoing the sperm formation sequence are positively correlated with the fluorescence patterns of the antibodies against SF53/4, SC35, and the Sm antigens. These data suggest that cessation of transcription during spermatogenesis is accompanied by exclusion of the splicing machinery from nontranscribing chromatin to its vicinity.
本文的主要目的是研究在一个大的核区域中,转录程序终止对前体mRNA剪接机制元件分布的影响。为此,我们利用了减数分裂早期小鼠精母细胞的核分区,即常染色体转录区和XY非转录区。该系统还允许将这项研究扩展到精子分化阶段,这些阶段伴随着转录的减少和最终终止。我们通过对生精细胞的间接免疫荧光显示:1)前体mRNA剪接因子SF53/4和SC35、Sm抗原以及RNA聚合酶II的荧光信号,在非转录的X失活区域基本不存在,但在转录的常染色体区域大量存在;2)在经历精子形成序列的细胞核中,转录活性的存在、逐渐降低和消失与针对SF53/4、SC35和Sm抗原的抗体的荧光模式呈正相关。这些数据表明,精子发生过程中转录的终止伴随着剪接机制从非转录染色质区域被排除到其附近区域。