Mobility of mPing and its associated elements is regulated by both internal and terminal sequences.

IF 4.7 2区 生物学 Q1 GENETICS & HEREDITY Mobile DNA Pub Date : 2023-02-11 DOI:10.1186/s13100-023-00289-3
Priscilla S Redd, Stephanie Diaz, David Weidner, Jazmine Benjamin, C Nathan Hancock
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引用次数: 3

Abstract

Background: DNA transposable elements are mobilized by a "cut and paste" mechanism catalyzed by the binding of one or more transposase proteins to terminal inverted repeats (TIRs) to form a transpositional complex. Study of the rice genome indicates that the mPing element has experienced a recent burst in transposition compared to the closely related Ping and Pong elements. A previously developed yeast transposition assay allowed us to probe the role of both internal and terminal sequences in the mobilization of these elements.

Results: We observed that mPing and a synthetic mPong element have significantly higher transposition efficiency than the related autonomous Ping and Pong elements. Systematic mutation of the internal sequences of both mPing and mPong identified multiple regions that promote or inhibit transposition. Simultaneous alteration of single bases on both mPing TIRs resulted in a significant reduction in transposition frequency, indicating that each base plays a role in efficient transposase binding. Testing chimeric mPing and mPong elements verified the important role of both the TIRs and internal regulatory regions. Previous experiments showed that the G at position 16, adjacent to the 5' TIR, allows mPing to have higher mobility. Alteration of the 16th and 17th base from mPing's 3' end or replacement of the 3' end with Pong 3' sequences significantly increased transposition frequency.

Conclusions: As the transposase proteins were consistent throughout this study, we conclude that the observed transposition differences are due to the element sequences. The presence of sub-optimal internal regions and TIR bases supports a model in which transposable elements self-limit their activity to prevent host damage and detection by host regulatory mechanisms. Knowing the role of the TIRs, adjacent sub-TIRs, and internal regulatory sequences allows for the creation of hyperactive elements.

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mPing及其相关元件的迁移受内部序列和末端序列的调控。
背景:DNA转座因子通过一个或多个转座酶蛋白与末端倒置重复序列(TIRs)结合催化的“剪切和粘贴”机制被动员起来,形成转座复合物。对水稻基因组的研究表明,与与之密切相关的Ping和Pong元件相比,mPing元件最近经历了一次爆发性的转位。先前开发的酵母转位试验使我们能够探测内部和末端序列在这些元件动员中的作用。结果:我们观察到mPing和合成的mPong元件的转位效率明显高于相关的自主Ping和Pong元件。mPing和mPong内部序列的系统突变发现了促进或抑制转位的多个区域。同时改变两个mPing TIRs上的单个碱基导致转座频率显著降低,表明每个碱基都在有效的转座酶结合中发挥作用。嵌合mPing和mPong元件的测试证实了tir和内部调控区域的重要作用。先前的实验表明,位置16的G与5' TIR相邻,使得mPing具有更高的迁移率。从mPing的3'端改变第16和第17个碱基或用Pong的3'序列替换3'端显著增加了转位频率。结论:由于转座酶蛋白在整个研究中是一致的,我们得出结论,观察到的转座差异是由于元件序列所致。次优内部区域和TIR碱基的存在支持了转座因子自我限制其活性以防止宿主损伤和被宿主调节机制检测的模型。了解tir、相邻子tir和内部调控序列的作用,可以创建多活性元件。
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来源期刊
Mobile DNA
Mobile DNA GENETICS & HEREDITY-
CiteScore
8.20
自引率
6.10%
发文量
26
审稿时长
11 weeks
期刊介绍: Mobile DNA is an online, peer-reviewed, open access journal that publishes articles providing novel insights into DNA rearrangements in all organisms, ranging from transposition and other types of recombination mechanisms to patterns and processes of mobile element and host genome evolution. In addition, the journal will consider articles on the utility of mobile genetic elements in biotechnological methods and protocols.
期刊最新文献
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