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家蚕端粒重复序列特异性逆转座子TRAS1和SART1的转录分析

Transcription analysis of the telomeric repeat-specific retrotransposons TRAS1 and SART1 of the silkworm Bombyx mori.

作者信息

Takahashi H, Fujiwara H

机构信息

Department of Biological Sciences, Graduate School of Science, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku,Tokyo 113-0033, Japan.

出版信息

Nucleic Acids Res. 1999 May 1;27(9):2015-21. doi: 10.1093/nar/27.9.2015.

Abstract

The telomeres of the silkworm Bombyx mori consist of (TTAGG)n repeats and harbor a large number of sequence-specific non-LTR retrotransposons such as TRAS1 and SART1. In order to ascertain if TRAS1 and SART1 are transcribed in vivo and if there is a novel transcription mechanism peculiar to the sequence-specific retrotransposons, we studied their transcription. We detected transcripts of TRAS1 and SART1 by northern hybridization in many tissues and the BmN4 cell line of the silkworm. 5'-Rapid amplification of cDNA ends analysis showed that transcription of both elements was initiated precisely from their own 5'-ends and that most of their genomic copies contained these initiation sites. TRAS1 contained an internal promoter and positively regulating elements in the +1/+581 nucleotides in its 2432 bp 5'-untranslated region (UTR). We could not, however, detect any promoter activity in the SART1 5'-UTR. This difference may be related to the fact that only TRAS1 contained an initiator-like element at its 5'-end. Placing 1-52 units of the telomeric repeat (TTAGG)n upstream of TRAS1 reduced transcription 5-fold. The evidence suggests that most of the TRAS1 genomic copies within the telomeric repeats are weakly transcribed in vivo.

摘要

家蚕的端粒由(TTAGG)n重复序列组成,并含有大量序列特异性非LTR逆转座子,如TRAS1和SART1。为了确定TRAS1和SART1在体内是否转录,以及是否存在序列特异性逆转座子特有的新型转录机制,我们对它们的转录进行了研究。我们通过Northern杂交在蚕的许多组织和BmN4细胞系中检测到了TRAS1和SART1的转录本。5'-cDNA末端快速扩增分析表明,这两种元件的转录均精确地从其自身的5'-末端起始,并且它们的大多数基因组拷贝都包含这些起始位点。TRAS1在其2432 bp的5'-非翻译区(UTR)的+1/+581核苷酸中含有一个内部启动子和正向调控元件。然而,我们在SART1的5'-UTR中未检测到任何启动子活性。这种差异可能与只有TRAS1在其5'-末端含有类似起始子元件这一事实有关。将1-52个端粒重复序列(TTAGG)n置于TRAS1上游可使转录降低5倍。证据表明,端粒重复序列内的大多数TRAS1基因组拷贝在体内转录较弱。

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