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首页> 外文期刊>Molecular and Cellular Biology >RNA binding by Sxl proteins in vitro and in vivo.
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RNA binding by Sxl proteins in vitro and in vivo.

机译：Sxl蛋白在体外和体内与RNA结合。

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Sxl has been proposed to regulate splicing of specific target genes by directly interacting with their pre-mRNAs. We have therefore examined the RNA-binding properties of Sxl protein in vitro and in vivo. Gel shift and UV cross-linking assays with a purified recombinant MBP-Sxl fusion protein demonstrated preferential binding to RNAs containing poly(U) tracts, and the protein footprinted over the poly(U) region. The protein did not appear to recognize either branch point or AG dinucleotide sequences, but an adenosine residue at the 5' end of the poly(U) tract enhanced binding severalfold. MBP-Sxl formed two shifted complexes on a tra regulated acceptor site RNA; the doubly shifted form may have been stabilized by protein-protein interactions. Consistent with its proposed role in pre-mRNA processing, in nuclear extracts Sxl was found in large ribonucleoprotein (RNP) complexes which sedimented significantly faster than bulk heterogeneous nuclear RNP and small nuclear RNPs. Anti-Sxl staining of polytene chromosomes showed Sxl protein at a number of chromosomal locations, among which was the Sxl locus itself. Sxl protein could also be targeted to a new chromosomal site carrying a transgene containing splicing regulatory sequences from the Sxl gene, following transcriptional induction. After prolonged heat shock, all Sxl protein was restricted to the heat-induced puff at the hs93D locus. In contrast, a presumptive small nuclear RNP protein was observed at several heat puffs following shock.
机译：已经提出Sx1通过与它们的前mRNA直接相互作用来调节特定靶基因的剪接。因此，我们已经在体外和体内检查了Sxl蛋白的RNA结合特性。用纯化的重组MBP-Sxl融合蛋白进行的凝胶位移和UV交联测定证明与包含多聚（U）片段的RNA优先结合，并且该蛋白质在多聚（U）区域上覆盖。该蛋白似乎不识别分支点或AG二核苷酸序列，但在多聚（U）道5'端的腺苷残基可增强结合几倍。 MBP-Sxl在tra调节的受体位点RNA上形成了两个移位的复合物。双移位形式可能已通过蛋白质-蛋白质相互作用而稳定。与其拟议的在mRNA加工中的作用一致，在大型核糖核蛋白（RNP）复合物中发现了Sxl，其沉降速度明显快于大块异质核RNP和小核RNP。多染色体染色体的抗Sxl染色在许多染色体位置显示Sxl蛋白，其中包括Sxl基因座本身。在转录诱导后，Sxl蛋白也可以靶向带有转基因的新染色体位点，该转基因包含来自Sxl基因的剪接调控序列。长时间的热休克后，所有的Sxl蛋白都被限制在hs93D基因座上的热诱导粉扑中。相反，在电击后的若干次热风抽吸中观察到推测的小核RNP蛋白。

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