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紫外线B照射诱导果蝇1731反转录转座子长末端重复序列上调需要U3区域的一段短序列。

Upregulation of the Drosophila 1731 retrotransposon long-terminal repeat by UV-B irradiation requires a short sequence in the U3 region.

作者信息

Faure E, Best-Belpomme M, Champion S

机构信息

Institut de Chimie Biologique, Université de Provence, Marseille, France.

出版信息

Arch Biochem Biophys. 1996 Feb 15;326(2):219-26. doi: 10.1006/abbi.1996.0068.

Abstract

A 1731 is a Drosophila melanogaster retrotransposon, the transcripts of which decrease in Drosophila cells after treatment by the 20-hydroxyecdysone (20-OH), the steroid-molting hormone of insects. In order to analyze the regulation of the long terminal repeat (LTR) directed transcription by UV-B, S2 Drosophila cells were transfected with various chimeric constructs carrying the LTR of 1731 linked to the bacterial chloramphenicol acetyltransferase (CAT) reporter gene and then subjected to UV-B irradiation. The results demonstrated that the 1731 LTR is activated by UV-B irradiation in a dose- and time-dependent manner. Using constructions expressing the reporter gene under the control of either the entire or deleted LTRs of 1731, we established that a sequence located in the U3 region was required for the retrotransposon to respond to UV-B. The cis-acting element is identical to the binding sequence of the dorsal transcription factors. In addition, the S2 Drosophila cell produced extracellular factor(s) in response to UV-B irradiation which activated the 1731 LTR in nonirradiated cells. This factor(s) was detected when responding cells were cocultured with inducing cells and when conditioned medium from irradiated cultures was added to the cell cultures.

摘要

A 1731是一种黑腹果蝇反转录转座子,在经昆虫类固醇蜕皮激素20-羟基蜕皮酮(20-OH)处理后的果蝇细胞中,其转录本会减少。为了分析UV-B对长末端重复序列(LTR)指导转录的调控作用,将携带与细菌氯霉素乙酰转移酶(CAT)报告基因相连的1731的LTR的各种嵌合构建体转染到S2果蝇细胞中,然后进行UV-B照射。结果表明,1731 LTR被UV-B照射以剂量和时间依赖性方式激活。利用在1731的完整或缺失LTR控制下表达报告基因的构建体,我们确定位于U3区域的一个序列是反转录转座子对UV-B作出反应所必需的。顺式作用元件与背侧转录因子的结合序列相同。此外,S2果蝇细胞在受到UV-B照射时会产生细胞外因子,该因子可激活未照射细胞中的1731 LTR。当反应细胞与诱导细胞共培养以及将照射培养物的条件培养基添加到细胞培养物中时,均可检测到这种因子。

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