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利用二分Alu逆转座法（BAR）鉴定对逆转座重要的L1 ORF2p序列。

Identification of L1 ORF2p sequence important to retrotransposition using Bipartile Alu retrotransposition (BAR).

作者信息

Christian Claiborne M, deHaro Dawn, Kines Kristine J, Sokolowski Mark, Belancio Victoria P

机构信息

Structural and Cellular Biology, Tulane University School of Medicine, Tulane Cancer Center, Tulane Center for Aging, New Orleans, LA 70130, USA.

Structural and Cellular Biology, Tulane University School of Medicine, Tulane Cancer Center, Tulane Center for Aging, New Orleans, LA 70130, USA

出版信息

Nucleic Acids Res. 2016 Jun 2;44(10):4818-34. doi: 10.1093/nar/gkw277. Epub 2016 Apr 19.

DOI:10.1093/nar/gkw277
PMID:27095191
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4889948/
Abstract

Long Interspersed Element 1 (LINE-1 or L1) is capable of causing genomic instability through the activity of the L1 ORF2 protein (ORF2p). This protein contains endonuclease (EN) and reverse transcriptase (RT) domains that are necessary for the retrotransposition of L1 and the Short Interspersed Element (SINE) Alu. The functional importance of approximately 50% of the ORF2p molecule remains unknown, but some of these sequences could play a role in retrotransposition, or be necessary for the enzymatic activities of the EN and/or RT domains. Conventional approaches using the full-length, contiguous ORF2p make it difficult to study the involvement of these unannotated sequences in the function of L1 ORF2p. Our lab has developed a Bipartile Alu Retrotransposition (BAR) assay that relies on separate truncated ORF2p fragments: an EN-containing and an RT-containing fragment. We validated the utility of this method for studying the ORF2p function in retrotransposition by assessing the effect of expression levels and previously characterized mutations on BAR. Using BAR, we identified two pairs of amino acids important for retrotransposition, an FF and a WD. The WD appears to play a role in cDNA synthesis by the ORF2p molecule, despite being outside the canonical RT domain.

摘要

长散在核元件1（LINE-1或L1）能够通过L1开放阅读框2蛋白（ORF2p）的活性导致基因组不稳定。该蛋白包含内切核酸酶（EN）和逆转录酶（RT）结构域，这些结构域对于L1和短散在核元件（SINE）Alu的逆转座是必需的。ORF2p分子约50%的功能重要性仍然未知，但其中一些序列可能在逆转座中发挥作用，或者对于EN和/或RT结构域的酶活性是必需的。使用全长、连续的ORF2p的传统方法难以研究这些未注释序列在L1 ORF2p功能中的参与情况。我们实验室开发了一种二分Alu逆转座（BAR）测定法，该方法依赖于单独的截短ORF2p片段：一个含EN的片段和一个含RT的片段。我们通过评估表达水平和先前表征的突变对BAR的影响，验证了该方法在研究ORF2p在逆转座中的功能方面的实用性。使用BAR，我们鉴定出两对对于逆转座很重要的氨基酸，一对FF和一对WD。尽管WD位于经典RT结构域之外，但它似乎在ORF2p分子的cDNA合成中发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1479/4889948/3f91670e71f3/gkw277fig17.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1479/4889948/3f91670e71f3/gkw277fig17.jpg

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1
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2
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Proc Natl Acad Sci U S A. 2015 Sep 1;112(35):E4894-900. doi: 10.1073/pnas.1502474112. Epub 2015 Aug 17.
3
The mechanism of ageing: primary role of transposable elements in genome disintegration.
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Int J Mol Sci. 2023 May 27;24(11):9380. doi: 10.3390/ijms24119380.
4
A novel role of TRIM28 B box domain in L1 retrotransposition and ORF2p-mediated cDNA synthesis.TRIM28 B 盒结构域在 L1 逆转录转座和 ORF2p 介导的 cDNA 合成中的新作用。
Nucleic Acids Res. 2023 May 22;51(9):4429-4450. doi: 10.1093/nar/gkad247.
5
The L1-dependant and Pol III transcribed Alu retrotransposon, from its discovery to innate immunity.从发现到先天免疫：L1 依赖性和 Pol III 转录的 Alu 反转录转座子。
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6
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Genetics. 2019 Dec;213(4):1401-1414. doi: 10.1534/genetics.119.302601. Epub 2019 Oct 30.
7
Truncated ORF1 proteins can suppress LINE-1 retrotransposition in trans.截短的ORF1蛋白可以反式抑制LINE-1逆转录转座。
Nucleic Acids Res. 2017 May 19;45(9):5294-5308. doi: 10.1093/nar/gkx211.
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9
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