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奇妙的复合结构,由反转录转座子和与美洲摇蚊(双翅目:摇蚊科)染色体末端相关的复杂重复序列组成。

Curiously composite structures of a retrotransposon and a complex repeat associated with chromosome ends of Rhynchosciara americana (Diptera: Sciaridae).

机构信息

Departamento de Genética e Biologia Evolutiva, Instituto de Biociências, Universidade de São Paulo, Rua do Matão 277, Cidade Universitária, São Paulo, SP, 05508-090, Brazil.

出版信息

Chromosome Res. 2010 Jul;18(5):587-98. doi: 10.1007/s10577-010-9143-x. Epub 2010 Jul 7.

Abstract

In Drosophila, telomere retrotransposons counterbalance the loss of telomeric DNA. The exceptional mechanism of telomere recovery characterized in Drosophila has not been found in lower dipterans (Nematocera). However, a retroelement resembling a telomere transposon and termed "RaTART" has been described in the nematoceran Rhynchosciara americana. In this work, DNA and protein sequence analyses, DNA cloning, and chromosomal localization of probes obtained either by PCR or by screening a genomic library were carried out in order to examine additional features of this retroelement. The analyses performed raise the possibility that RaTART represents a genomic clone composed of distinct repetitive elements, one of which is likely to be responsible for its apparent enrichment at chromosome ends. RaTART sequence in addition allowed to assess a novel subtelomeric region of R. americana chromosomes that was analyzed in this work after subcloning a DNA fragment from a phage insert. It contains a complex repeat that is located in the vicinity of simple and complex tandem repeats characterized previously. Quantification data suggest that the copy number of the repeat is significantly lower than that observed for the ribosomal DNA in the salivary gland of R. americana. A short insertion of the RaTART was identified in the cloned segment, which hybridized preferentially to subtelomeres. Like RaTART, it displays truncated sequences related to distinct retrotransposons, one of which has a conceptual translation product with significant identity with an endonuclease from a lepidopteran retrotransposon. The composite structure of this DNA stretch probably reflects mobile element activity in the subtelomeric region analyzed in this work.

摘要

在果蝇中,端粒反转录转座子平衡了端粒 DNA 的损失。在较低等的双翅目昆虫(Nematocera)中尚未发现这种在果蝇中具有特征的端粒恢复的特殊机制。然而,在 Nematoceran Rhynchosciara americana 中已经描述了一个类似于端粒转座子的反转录元件,称为“RaTART”。在这项工作中,进行了 DNA 和蛋白质序列分析、DNA 克隆以及通过 PCR 或筛选基因组文库获得的探针的染色体定位,以检查该反转录元件的其他特征。进行的分析提出了这样的可能性,即 RaTART 代表由不同重复元件组成的基因组克隆,其中一个元件可能负责其在染色体末端的明显富集。RaTART 序列还允许评估 R. americana 染色体的一个新的亚端粒区域,在这项工作中,在从噬菌体插入物中克隆一个 DNA 片段后,对其进行了分析。它包含一个复杂的重复序列,位于先前描述的简单和复杂串联重复序列的附近。定量数据表明,该重复序列的拷贝数明显低于 R. americana 唾液腺中核糖体 DNA 的观察值。在克隆片段中鉴定出 RaTART 的一个短插入,它优先与亚端粒杂交。与 RaTART 一样,它显示出与不同反转录转座子相关的截断序列,其中一个具有概念性翻译产物,与鳞翅目反转录转座子的内切核酸酶具有显著的同一性。这段 DNA 的复合结构可能反映了在这项工作中分析的亚端粒区域中移动元件的活性。

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