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家蚕中新型端粒重复序列相关反转录转座子家族TRAS1的结构分析

Structural analysis of TRAS1, a novel family of telomeric repeat-associated retrotransposons in the silkworm, Bombyx mori.

作者信息

Okazaki S, Ishikawa H, Fujiwara H

机构信息

Zoological Institute, Graduate School of Science, University of Tokyo, Japan.

出版信息

Mol Cell Biol. 1995 Aug;15(8):4545-52. doi: 10.1128/MCB.15.8.4545.

Abstract

We characterized TRAS1, a retrotransposable element which was inserted into the telomeric repetitive sequence (CCTAA)n of the silkworm, Bombyx mori. The complete sequence of TRAS1, a stretch of 7.8 kb with a poly(A) tract at the 3' end, was determined. No long terminal repeat (LTR) was found at the termini of the element. TRAS1 contains gag- and pol-like open reading frames (ORFs) which are similar to those of non-LTR retrotransposons. The two ORFs overlap but are one nucleotide out of frame (+1 frameshift). Most of the approximately 250 copies of TRAS1 elements in the genome were highly conserved in the structure. Chromosomal in situ hybridization showed that TRAS1 elements are clustered at the telomeres of Bombyx chromosomes. A phylogenetic analysis using the amino acid sequence of the reverse transcriptase domain within the pol-like ORF revealed that TRAS1 falls into one lineage with R1, which is a family of non-LTR retrotransposons inserted into the same site within the 28S ribosomal DNA unit in most insects. TRAS1 may have been derived from R1 and changed the target specificity so that TRAS1 inserts into the telomeric repetitive sequence (CCTAA)n. Southern hybridization and Bal 31 exonuclease analyses showed that TRAS1 elements are clustered proximal to the terminal long tract of (CCTAA)n. TRAS1 is a novel family of non-LTR retrotransposons which are inserted into the telomeric repetitive sequences as target sites.

摘要

我们对TRAS1进行了特征分析,它是一种逆转座子元件,插入到家蚕(Bombyx mori)的端粒重复序列(CCTAA)n中。确定了TRAS1的完整序列,其长度为7.8 kb,3'端有一个聚腺苷酸尾。在该元件的末端未发现长末端重复序列(LTR)。TRAS1包含与非LTR逆转座子相似的gag和pol样开放阅读框(ORF)。这两个ORF重叠,但有一个核苷酸移码(+1移码)。基因组中大约250个TRAS1元件的大部分在结构上高度保守。染色体原位杂交显示TRAS1元件聚集在家蚕染色体的端粒处。使用pol样ORF内逆转录酶结构域的氨基酸序列进行的系统发育分析表明,TRAS1与R1属于同一谱系,R1是一类非LTR逆转座子家族,在大多数昆虫的28S核糖体DNA单元内插入到相同位点。TRAS1可能源自R1并改变了靶标特异性,从而使TRAS1插入端粒重复序列(CCTAA)n。Southern杂交和Bal 31核酸外切酶分析表明,TRAS1元件聚集在(CCTAA)n末端长序列的近端。TRAS1是一类新型的非LTR逆转座子,以端粒重复序列作为靶位点进行插入。

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