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首页> 外文期刊>Molecular and Cellular Biology >The concentration of B52, an essential splicing factor and regulator of splice site choice in vitro, is critical for Drosophila development.
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The concentration of B52, an essential splicing factor and regulator of splice site choice in vitro, is critical for Drosophila development.

机译：B52的浓度是必需的剪接因子和体外剪接位点选择的调节剂，对果蝇的发育至关重要。

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B52 is a Drosophila melanogaster protein that plays a role in general and alternative splicing in vitro. It is homologous to the human splicing factor ASF/SF2 which is essential for an early step(s) in spliceosome assembly in vitro and also regulates 5' and 3' alternative splice site choice in a concentration-dependent manner. In vitro, B52 can function as both a general splicing factor and a regulator of 5' alternative splice site choice. Its activity in vivo, however, is largely uncharacterized. In this study, we have further characterized B52 in vivo. Using Western blot (immunoblot) analysis and whole-mount immunofluorescence, we demonstrate that B52 is widely expressed throughout development, although some developmental stages and tissues appear to have higher B52 levels than others do. In particular, B52 accumulates in ovaries, where it is packaged into the developing egg and is localized to nuclei by the late blastoderm stage of embryonic development. We also overexpressed this protein in transgenic flies in a variety of developmental and tissue-specific patterns to examine the effects of altering the concentration of this splicing factor in vivo. We show that, in many cell types, changing the concentration of B52 adversely affects the development of the organism. We discuss the significance of these observations with regard to previous in vitro results.
机译：B52是果蝇的果蝇蛋白，在体外普通剪接过程中发挥作用。它与人剪接因子ASF / SF2同源，后者对于体外剪接体组装的早期步骤是必不可少的，并且还以浓度依赖的方式调节5'和3'替代剪接位点的选择。在体外，B52既可作为一般剪接因子，又可作为5'替代剪接位点选择的调节剂。然而，其在体内的活性在很大程度上未被表征。在这项研究中，我们进一步表征了体内的B52。使用蛋白质印迹（免疫印迹）分析和全定量免疫荧光，我们证明了B52在整个发育过程中广泛表达，尽管某些发育阶段和组织似乎比其他发育阶段具有更高的B52水平。特别地，B52积累在卵巢中，然后被包装到发育中的卵中，并在胚胎发育的胚盘末期定位到细胞核中。我们还以各种发育和组织特异性模式在转基因果蝇中过表达了这种蛋白质，以研究改变这种剪接因子在体内浓度的影响。我们表明，在许多细胞类型中，改变B52的浓度会对有机体的发育产生不利影响。我们讨论有关以前的体外结果这些意见的意义。

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