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首页> 外文期刊>Molecular and Cellular Biology >The Drosophila suppressor of sable protein binds to RNA and associates with a subset of polytene chromosome bands.
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The Drosophila suppressor of sable protein binds to RNA and associates with a subset of polytene chromosome bands.

机译：果蝇蛋白的果蝇抑制剂与RNA结合并与多聚体染色体带子集相关。

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Mutations of the Drosophila melanogaster suppressor of sable [su(s)] gene, which encodes a 150-kDa nuclear protein [Su(s)], increase the accumulation of specific transcripts in a manner that is not well understood but that appears to involve pre-mRNA processing. Here, we report biochemical analysis of purified, recombinant Su(s) [rSu(s)] expressed in baculovirus and in Escherichia coli as maltose binding protein (MBP) fusions and immunocytochemical analysis of endogenous Su(s). This work has shown that purified, baculovirus-expressed rSu(s) binds to RNA in vitro with a high affinity and limited specificity. Systematic evolution of ligands by exponential enrichment was used to identify preferred RNA targets of rSu(s), and a large proportion of RNAs isolated contain a full or partial match to the consensus sequence UCAGUAGUCU, which was confirmed to be a high-affinity rSu(s) binding site. An MBP-Su(s) fusion protein containing the N-terminal third of Su(s) binds RNAs containing this sequence with a higher specificity than full-length, baculovirus-expressed rSu(s). The consensus sequence resembles both a cryptic 5' splice site and a sequence that is found near the 5' end of some Drosophila transcripts. Immunolocalization studies showed that endogenous Su(s) is distributed in a reticulated pattern in Drosophila embryo and salivary gland nuclei. In salivary gland cells, Su(s) is found both in the nucleoplasm and in association with a subset of polytene chromosome bands. Considering these and previous results, we propose two models to explain how su(s) mutations affect nuclear pre-mRNA processing.
机译：果蝇[su（s）]基因的果蝇黑色素抑制因子的突变，其编码150 kDa核蛋白[Su（s）]，以一种尚不十分了解但似乎涉及的方式增加了特定转录本的积累。 mRNA前处理。在这里，我们报告了在杆状病毒和大肠杆菌中表达为麦芽糖结合蛋白（MBP）融合蛋白的纯化，重组Su [rSu（s）]的生化分析和内源Su（s）的免疫细胞化学分析。这项工作表明，纯化的杆状病毒表达的rSu在体外以高亲和力和有限的特异性与RNA结合。配体通过指数富集的系统进化被用于识别rSu的首选RNA靶标，并且分离出的大部分RNA包含与共有序列UCAGUAGUCU的全部或部分匹配，这被证实是高亲和性rSu（ s）结合位点。包含Su的N末端三分之一的MBP-Su融合蛋白以比全长杆状病毒表达的rSu更高的特异性结合含有该序列的RNA。共有序列既类似于隐蔽的5'剪接位点，又类似于在某些果蝇转录本的5'末端附近发现的序列。免疫定位研究表明，内源性Su（s）以网状分布在果蝇胚胎和唾液腺核中。在唾液腺细胞中，在核质中以及与多聚体染色体带的子集相关的地方都发现了Su。考虑到这些和以前的结果，我们提出了两个模型来解释su（s）突变如何影响mRNA前核的加工。

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