The C-terminus of the Hermes transposase contains a protein multimerization domain.
The C-terminus of the Hermes transposase contains a protein multimerization domain.
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DOI：
10.1016/s0965-1748(03)00102-4
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发表时间：
2003-10
影响因子：
3.8
通讯作者：
K. Michel;D. O’brochta;P. Atkinson
中科院分区：
文献类型：
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作者：
K. Michel;D. O’brochta;P. Atkinson
介导 II 类转座元件移动性的转座酶活性通常是由称为转座体的高级突触复合体的组装引发的。这些复合物包含转座元件的 DNA 以及两个或多个转座酶分子，其形成取决于转座酶分子之间的相互作用。使用酵母双杂交系统，我们能够识别介导 Hermes 转座酶多聚化的三个区域，Hermes 转座酶是一种用于昆虫种系转化的元件，属于 hAT 转座元件家族。在转座酶的前 252 个氨基酸内发现了一个促进 Hermes 转座酶分子蛋白质结合的区域。第二个区域位于转座酶的 C 末端，被发现对 Hermes 转座酶多聚化具有特异性。氨基酸 551-569 不仅是多聚化所必需的，而且也是元件转座所必需的。第三个区域位于氨基酸 253 和 380 之间，被发现可以消除 N 端蛋白质相互作用区域的非特异性蛋白质结合能力，但对于转座酶 C 端区域的特异性蛋白质结合能力来说是必需的。影响 C 端多聚化区域结构完整性的五个点突变消除或显着降低了转座活性。先前已鉴定出同一区域介导另一种 hAT 元件激活剂 (Ac) 的二聚化，表明 hAT 转座酶多聚化可能是其元件移动性的先决条件。
Transposase activity that mediates the mobility of class II transposable elements, is most commonly initiated by the assembly of higher order synaptic complexes, called transpososomes. The formation of these complexes, that contain the transposable element’s DNA as well as two or more molecules of the transposase, is dependent on interactions between transposase molecules. Using the yeast Two-Hybrid system, we were able to identify three regions mediating multimerization of the Hermes transposase, an element used for germline transformation of insects belonging to the hAT family of transposable elements. One region facilitating protein binding of Hermes transposase molecules was found within the first 252 amino acids of the transposase. The second region was located at the C-terminus of the transposase, and was found to be specific for Hermes transposase multimerization. Amino acids 551–569 were not only required for multimerization but were also necessary for transposition of the element. The third region was located between amino acids 253 and 380 and was found to eliminate the non-specific protein binding ability of the N-terminal protein interaction region but was required for the specific protein binding ability of the C-terminal region of the transposase. Five point mutations affecting the structural integrity of the C-terminal multimerization region abolished or significantly reduced transpositional activity. The same region had been previously identified to mediate dimerization in Activator (Ac), another hAT element, indicating that hAT transposase multimerization is likely to be a prerequisite for mobility of their elements.