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鸡β-H珠蛋白基因处的转录终止

Transcription termination at the chicken beta H-globin gene.

作者信息

Pribyl T M, Martinson H G

机构信息

Department of Chemistry and Biochemistry, University of California, Los Angeles 90024.

出版信息

Mol Cell Biol. 1988 Dec;8(12):5369-77. doi: 10.1128/mcb.8.12.5369-5377.1988.

Abstract

We characterized the transcription termination region of the chicken beta H-globin gene. First we located the region by nuclear runon transcription in vitro. Then we sequenced and subcloned it into a chloramphenicol acetyltransferase (CAT) expression vector for assay in vivo. The region of beta H termination contains two interesting elements located about 1 kilobase downstream of the beta H gene poly(A) site. Either element alone can block CAT expression if inserted between the promoter and the poly(A) site of the cat gene in pRSVcat. The first element in the termination region is an unusually large inverted repeat in the DNA (delta G = -71 kcal). The second element, 200 base pairs further downstream, is an RNA polymerase II promoter which directs transcription back upstream on the complementary strand. This transcription converges on and collides with that from the beta H gene at or near the inverted repeat where transcription from both directions stops. We propose that the inverted repeat is a strong pause site which positions the converging polymerases for mutual site-specific termination.

摘要

我们对鸡βH-珠蛋白基因的转录终止区域进行了特征分析。首先,我们通过体外核转录延伸定位了该区域。然后,我们对其进行测序并亚克隆到氯霉素乙酰转移酶(CAT)表达载体中,以便在体内进行检测。βH终止区域包含两个有趣的元件,位于βH基因多聚腺苷酸化位点下游约1千碱基处。如果将这两个元件中的任何一个插入到pRSVcat中cat基因的启动子和多聚腺苷酸化位点之间,都能阻断CAT的表达。终止区域中的第一个元件是DNA中一个异常大的反向重复序列(ΔG = -71千卡)。第二个元件位于下游200个碱基对处,是一个RNA聚合酶II启动子,它在互补链上引导转录向上游进行。这种转录在反向重复序列处或其附近与来自βH基因的转录汇聚并发生碰撞,双向转录在此处停止。我们提出,反向重复序列是一个强暂停位点,它使汇聚的聚合酶定位,以进行相互的位点特异性终止。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c3a4/365639/12b38d111b65/molcellb00072-0323-a.jpg

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