Plant-exclusive domain of trans-editing enzyme ProXp-ala confers dimerization and enhanced tRNA binding.
Plant-exclusive domain of trans-editing enzyme ProXp-ala confers dimerization and enhanced tRNA binding.
复制标题
DOI：
10.1016/j.jbc.2022.102255
复制
发表时间：
2022-09
影响因子：
4.8
通讯作者：
Musier-Forsyth, Karin
中科院分区：
文献类型：
--
作者：
Byun, Jun-Kyu;Vu, John A.;He, Siou-Luan;Jang, Jyan-Chyun;Musier-Forsyth, Karin
Faithful translation of the genetic code is critical for the viability of all living organisms. The trans-editing enzyme ProXp-ala prevents Pro to Ala mutations during translation by hydrolyzing misacylated Ala-tRNAPro that has been synthesized by prolyl-tRNA synthetase. Plant ProXp-ala sequences contain a conserved C-terminal domain (CTD) that is absent in other organisms; the origin, structure, and function of this extra domain are unknown. To characterize the plant-specific CTD, we performed bioinformatics and computational analyses that provided a model consistent with a conserved α-helical structure. We also expressed and purified wildtype Arabidopsis thaliana (At) ProXp-ala in Escherichia coli, as well as variants lacking the CTD or containing only the CTD. Circular dichroism spectroscopy confirmed a loss of α-helical signal intensity upon CTD truncation. Size-exclusion chromatography with multiangle laser-light scattering revealed that wildtype At ProXp-ala was primarily dimeric and CTD truncation abolished dimerization in vitro. Furthermore, bimolecular fluorescence complementation assays in At protoplasts support a role for the CTD in homodimerization in vivo. The deacylation rate of Ala-tRNAPro by At ProXp-ala was also significantly reduced in the absence of the CTD, and kinetic assays indicated that the reduction in activity is primarily due to a tRNA binding defect. Overall, these results broaden our understanding of eukaryotic translational fidelity in the plant kingdom. Our study reveals that the plant-specific CTD plays a significant role in substrate binding and canonical editing function. Through its ability to facilitate protein–protein interactions, we propose the CTD may also provide expanded functional potential for trans-editing enzymes in plants.
登录
查看更多内容
影响因子：
14.9
作者：
Chan PP;Lowe TM
通讯作者：
Lowe TM
在古细胞RNase P中解剖蛋白质亚基（一种催化核糖核蛋白复合物）中的功能合作。
影响因子：
14.9
作者：
Chen WY;Pulukkunat DK;Cho IM;Tsai HY;Gopalan V
通讯作者：
Gopalan V
影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
影响因子：
3.5
作者：
Saga Y;Kawashima M;Sakai S;Yamazaki K;Kaneko M;Takahashi M;Sato N;Toyoda Y;Takase S;Nakano T;Kawakami N;Kushiro T
通讯作者：
Kushiro T
影响因子：
4.8
作者：
Cho, Ha Yeon;Maeng, Seo Jin;Kim, Sunghoon
通讯作者：
Kim, Sunghoon