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内含子反义RNA和核糖体蛋白S14对人RPS14转录的调控

Regulation of human RPS14 transcription by intronic antisense RNAs and ribosomal protein S14.

作者信息

Tasheva E S, Roufa D J

机构信息

Division of Biology, Kansas State University, Manhattan 66506-4901.

出版信息

Genes Dev. 1995 Feb 1;9(3):304-16. doi: 10.1101/gad.9.3.304.

Abstract

RNase protection studies reveal two stable RNAs (250 and 280 nucleotides) transcribed from the antisense strand of the human ribosomal protein gene RPS14's first intron. These transcripts, designated alpha-250 and alpha-280, map to overlapping segments of the intron's 5' sequence. Neither RNA encodes a polypeptide sequence, and both are expressed in all human cells and tissues examined. Although alpha-280 is detected among both the cells' nuclear and cytoplasmic RNAs, the great majority of alpha-250 is found in the cytoplasmic subcellular compartment. As judged by its resistance to high concentrations of alpha-amanitin, cell-free transcription of alpha-250 and alpha-280 appears to involve RNA polymerase I. Tissue culture transfection and cell-free transcription experiments demonstrate that alpha-250 and alpha-280 stimulate S14 mRNA transcription, whereas free ribosomal protein S14 inhibits it. Electrophoretic mobility shift experiments indicate specific binary molecular interactions between r-protein S14, its message and the antisense RNAs. In light of these data, we propose a model for fine regulation of human RPS14 transcription that involves RPS14 intron 1 antisense RNAs as positive effectors and S14 protein as a negative effector.

摘要

核糖核酸酶保护实验揭示,从人类核糖体蛋白基因RPS14第一个内含子的反义链转录出两种稳定的RNA(分别为250和280个核苷酸)。这些转录本,命名为α-250和α-280,定位于内含子5'序列的重叠片段。这两种RNA均不编码多肽序列,且在所检测的所有人类细胞和组织中均有表达。虽然在细胞核和细胞质RNA中均能检测到α-280,但绝大多数α-250存在于细胞质亚细胞区室中。根据其对高浓度α-鹅膏蕈碱的抗性判断,α-250和α-280的无细胞转录似乎涉及RNA聚合酶I。组织培养转染和无细胞转录实验表明,α-250和α-280可刺激S14 mRNA转录,而游离的核糖体蛋白S14则抑制该转录。电泳迁移率变动实验表明,核糖体蛋白S14、其信使RNA与反义RNA之间存在特定的二元分子相互作用。鉴于这些数据,我们提出了一个人类RPS14转录精细调控模型,该模型涉及将RPS14内含子1反义RNA作为正效应物,将S14蛋白作为负效应物。

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