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小鼠中移动性ETnII逆转录元件及其具有编码能力的MusD相关元件的结构与表达

Structure and expression of mobile ETnII retroelements and their coding-competent MusD relatives in the mouse.

作者信息

Baust Corinna, Gagnier Liane, Baillie Greg J, Harris Muriel J, Juriloff Diana M, Mager Dixie L

机构信息

Terry Fox Laboratory, B. C. Cancer Agency, Department of Medical Genetics, University of British Columbia, Vancouver, British Columbia, Canada.

出版信息

J Virol. 2003 Nov;77(21):11448-58. doi: 10.1128/jvi.77.21.11448-11458.2003.

Abstract

ETnII elements are mobile members of the repetitive early transposon family of mouse long terminal repeat (LTR) retroelements and have caused a number of mutations by inserting into genes. ETnII sequences lack retroviral genes, but the recent discovery of related MusD retroviral elements with regions similar to gag, pro, and pol suggests that MusD provides the proteins necessary for ETnII transposition in trans. For this study, we analyzed all ETnII elements in the draft sequence of the C57BL/6J genome and classified them into three subtypes (alpha, beta, and gamma) based on structural differences. We then used database searches and quantitative real-time PCR to determine the copy number and expression of ETnII and MusD elements in various mouse strains. In 7.5-day-old embryos of a mouse strain in which two mutations due to ETnII-beta insertions have been identified (SELH/Bc), we detected a three- to sixfold higher level of ETnII-beta and MusD transcripts than in control strains (C57BL/6J and LM/Bc). The increased ETnII transcription level can in part be attributed to a higher number of ETnII-beta elements, but 70% of the MusD transcripts appear to have been derived from one or a few MusD elements that are not detectable in C57BL/6J mice. This element belongs to a young MusD subgroup with intact open reading frames and identical LTRs, suggesting that the overexpressed element(s) in SELH/Bc mice might provide the proteins for the retrotransposition of ETnII and MusD elements. We also show that ETnII is expressed up to 30-fold more than MusD, which could explain why only ETnII, but not MusD, elements have been positively identified as new insertions.

摘要

ETnII元件是小鼠长末端重复序列(LTR)逆转录元件中重复早期转座子家族的可移动成员,通过插入基因已导致了许多突变。ETnII序列缺乏逆转录病毒基因,但最近发现的相关MusD逆转录病毒元件具有与gag、pro和pol相似的区域,这表明MusD提供了ETnII反式转座所需的蛋白质。在本研究中,我们分析了C57BL/6J基因组草图序列中的所有ETnII元件,并根据结构差异将它们分为三个亚型(α、β和γ)。然后,我们使用数据库搜索和定量实时PCR来确定各种小鼠品系中ETnII和MusD元件的拷贝数和表达情况。在一个已鉴定出由于ETnII-β插入导致两个突变的小鼠品系(SELH/Bc)的7.5天大的胚胎中,我们检测到ETnII-β和MusD转录本水平比对照品系(C57BL/6J和LM/Bc)高3至6倍。ETnII转录水平的升高部分可归因于ETnII-β元件数量的增加,但70%的MusD转录本似乎来自C57BL/6J小鼠中无法检测到的一个或几个MusD元件。该元件属于一个具有完整开放阅读框和相同LTR的年轻MusD亚组,这表明SELH/Bc小鼠中过表达的元件可能为ETnII和MusD元件的逆转录转座提供蛋白质。我们还表明,ETnII的表达比MusD高30倍,这可以解释为什么只有ETnII元件,而不是MusD元件,被确认为新的插入元件。

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