人α样珠蛋白基因簇中Alu家族RNA聚合酶III转录单位的组织、结构和体外转录:狼疮抗la抗体体外转录物的沉淀

C K Shen, T Maniatis
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摘要

我们研究了人类α样珠蛋白基因簇中重复DNA序列的位置、结构和体外转录。至少鉴定出8个不同的Alu家族重复序列,每个重复序列在体外转录产生离散的RNA转录物。测定了位于α 1珠蛋白基因3'侧(3'- α 1)的一个Alu重复序列的核苷酸序列,并与已发表的Alu重复序列进行了比较。该重复序列的体外转录产生大约410、260、160和86个核苷酸的RNA片段。为了确定这些转录本是否与体外特异性蛋白相关,我们使用系统性红斑狼疮(SLE)患者的抗血清进行了免疫沉淀实验。我们发现,抗血清anti-La可以从病毒感染的细胞中沉淀含有腺病毒VAI RNA的核糖核蛋白(RNPs),优先沉淀最小的两个体外3'- α 1 Alu重复序列的转录本。这些结果表明,rna与体外转录反应混合物中的特定因子相互作用形成RNP。
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The organization, structure, and in vitro transcription of Alu family RNA polymerase III transcription units in the human alpha-like globin gene cluster: precipitation of in vitro transcripts by lupus anti-La antibodies.

We have studied the location, structure, and in vitro transcription of repetitive DNA sequences within the human alpha-like globin gene cluster. At least eight different Alu family repeats were identified, each of which is transcribed in vitro to produce discrete RNA transcripts. The nucleotide sequence of one Alu repeat sequence, located on the 3' side of the alpha l globin gene (3'-alpha l), was determined and compared to published Alu repeat sequences. In vitro transcription of this repeat sequence generates RNA fragments of approximately 410, 260, 160, and 86 nucleotides. To determine whether these transcripts associate with specific proteins in vitro, we carried out immunoprecipitation experiments using an antiserum from systemic lupus erythematosus (SLE) patients. We find that the antiserum anti-La, which was shown to precipitate ribonucleoproteins (RNPs) containing the adenovirus VAI RNA from virus infected cells, preferentially precipitates the smallest two in vitro transcripts of the 3'-alpha l Alu repeat. These results suggest that the RNAs interact with specific factors in the in vitro transcription reaction mix to form RNP.

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