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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >RNA degradosomes exist in vivo in Escherichia coli as multicomponent complexes associated with the cytoplasmic membrane via the N-terminal region of ribonuclease E
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RNA degradosomes exist in vivo in Escherichia coli as multicomponent complexes associated with the cytoplasmic membrane via the N-terminal region of ribonuclease E

机译：RNA降解体作为通过核糖核酸酶E N端区域与细胞质膜结合的多组分复合物存在于大肠杆菌中

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摘要

RNase E isolated from Escherichia coli is contained in a multicom- ponent "degradosome" complex with other proteins implicated in RNA decay. Earlier work has shown that the C-terminal region of RNase E is a scaffold for the binding of degradosome components and has identified specific RNase E segments necessary for its interaction with polynucleotide phosphorylase (PNPase). RhlB RNA helicase, and enolase. Here, we report electron microscopy studies that use immunogold Labeling and freeze--fracture methods to show that degradosomes exist in vivo in E coli as multicomponent structures that associate with the cytoplasmic membrane via the N-terminal region of RNase E. Whereas PNPase and enolase are present in E coli in large excess relative to RNase E and therefore are detected in cells largely as molecules unlinked to the RNase E scaffold, immunogold labeling and biochemical analyses show that helicase is present in approximately equimolar amounts to RNase E at all cell groWth stages. Our findings. which establish the existence and cellular location of RNase E-based degradosomes in vivo in E. coli, also Suggest that RNA processing and decay may occur at specific sites within cells.
机译：从大肠杆菌中分离出的RNase E与其他与RNA降解有关的蛋白质一起包含在多组分“降解体”复合物中。较早的工作表明，RNase E的C端区域是结合降解体组分的支架，并鉴定了与多核苷酸磷酸化酶（PNPase）相互作用所必需的特定RNase E片段。 RhlB RNA解旋酶和烯醇化酶。在这里，我们报告了使用免疫金标记和冷冻断裂方法的电子显微镜研究，结果表明降解体以多组分结构存在于大肠杆菌中，并通过RNase E的N端区域与细胞质膜结合.PNPase和烯醇化酶大肠杆菌中相对于RNase E过量存在，因此在细胞中被大量检测为与RNase E支架未连接的分子，免疫金标记和生化分析表明解旋酶在所有细胞生长阶段均以与RNase E大约等摩尔的量存在。我们的发现。它们确定了体内基于RNase E的降解体在大肠杆菌中的存在和细胞位置，也表明RNA加工和衰变可能发生在细胞内的特定位点。

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