Hyperactive mariner transposons are created by mutations that disrupt allosterism and increase the rate of transposon end synapsis.
Hyperactive mariner transposons are created by mutations that disrupt allosterism and increase the rate of transposon end synapsis.
复制标题
DOI：
10.1093/nar/gkt1218
复制
发表时间：
2014-02
影响因子：
14.9
通讯作者：
Chalmers R
中科院分区：
文献类型：
--
作者：
Liu D;Chalmers R
转座子在脊椎动物遗传学中的新应用刺激了开发过度活跃变体的努力。通常，遗传筛选用于鉴定几个过度活跃的点突变，然后将其并入单个转座酶基因中。然而，负责增加活动的机制尚不清楚。在这里，我们表明，水手转座酶中的几个点突变增加其活动，破坏变构，通常用于下调转座通过减缓突触的转座子结束。我们专注于保守的WVPHEL氨基酸基序，它形成了水手转座酶二聚体界面的一部分。我们生成了W、V、E和L残基的几乎所有可能的单取代，并发现大多数是超活性的。对突变的生化分析显示，它们破坏了响应转座子末端结合而在发育中的转座体的相对侧之间传递的信号。除了它们在变构中的作用外，信号还控制催化的起始，从而防止非生产性双链断裂。最后，我们注意到，这种断裂可能解释了果蝇Mos1转座子末端令人困惑的“自我伤害”。
New applications for transposons in vertebrate genetics have spurred efforts to develop hyperactive variants. Typically, a genetic screen is used to identify several hyperactive point mutations, which are then incorporated in a single transposase gene. However, the mechanisms responsible for the increased activity are unknown. Here we show that several point mutations in the mariner transposase increase their activities by disrupting the allostery that normally serves to downregulate transposition by slowing synapsis of the transposon ends. We focused on the conserved WVPHEL amino acid motif, which forms part of the mariner transposase dimer interface. We generated almost all possible single substitutions of the W, V, E and L residues and found that the majority are hyperactive. Biochemical analysis of the mutations revealed that they disrupt signals that pass between opposite sides of the developing transpososome in response to transposon end binding. In addition to their role in allostery, the signals control the initiation of catalysis, thereby preventing non-productive double-strand breaks. Finally, we note that such breaks may explain the puzzling ‘self-inflicted wounds’ at the ends of the Mos1 transposon in Drosophila.
登录
查看更多内容
影响因子：
48
作者：
Yusa, Kosuke;Rad, Roland;Takeda, Junji;Bradley, Allan
通讯作者：
Bradley, Allan
影响因子：
3.5
作者：
Palomeque, T;Carrillo, JA;Lorite, P
通讯作者：
Lorite, P
影响因子：
14.9
作者：
Cuypers, Maxime G.;Trubitsyna, Maryia;Richardson, Julia M.
通讯作者：
Richardson, Julia M.
DOI：
10.1093/bfgp/2.1.57
发表时间：
2003-04-01
期刊：
Briefings in Functional Genomics & Proteomics
影响因子：
--
作者：
Ryder, Edward;Russell, Steven
通讯作者：
Russell, Steven
DOI：
10.1073/pnas.1008322108
发表时间：
2011-01-25
影响因子：
11.1
作者：
Yusa, Kosuke;Zhou, Liqin;Craig, Nancy L.
通讯作者：
Craig, Nancy L.