Use of a Yeast tRNase Killer Toxin to Diagnose Kti12 Motifs Required for tRNA Modification by Elongator.
Use of a Yeast tRNase Killer Toxin to Diagnose Kti12 Motifs Required for tRNA Modification by Elongator.
复制标题
DOI：
10.3390/toxins9090272
复制
发表时间：
2017-09-05
期刊：
影响因子：
4.2
通讯作者：
Schaffrath R
中科院分区：
文献类型：
--
作者：
Mehlgarten C;Prochaska H;Hammermeister A;Abdel-Fattah W;Wagner M;Krutyhołowa R;Jun SE;Kim GT;Glatt S;Breunig KD;Stark MJR;Schaffrath R
酿酒酵母细胞被来自乳酸克鲁维酵母（Kluyveromyceslactis）的tRNase核糖核酸毒素复合物（其切割反密码子并抑制蛋白质合成）的发酵素杀死。发酵霉素的作用需要通过延长子复合物（Elp 1-Elp 6）对反密码子摆动位置（U34）的尿苷碱基进行特异性化学修饰。因此，在缺乏Elongator或相关KTI的突变体中，反密码子修饰的丧失（K.乳酸毒素不敏感）基因保护免受tRNA切割并赋予对毒素的抗性。在这里，我们表明，酵母蛋白可以作为一种工具来遗传分析KTI 12，一个基因先前显示的编码的延伸伙伴蛋白。从一个kti 12突变池的zymocin幸存者，我们确定的基序在Kti 12功能直接耦合到延长器的活动。此外，无义和错义tRNA抑制（SUP 4; SOE 1）共同需要U34修饰，这强烈表明Kti 12和Elongator合作以确保正确的tRNA功能。我们发现Kti 12基序在来自拟南芥的植物直向同源物DRL 1/DRL 4中是保守的，并且似乎参与辅因子的结合（例如，核苷酸、钙调素）。由这些基序中的突变触发的延伸子相互作用缺陷与U34修饰缺失的典型表型相关。因此，由Elongator进行的tRNA修饰似乎需要与Kti 12进行物理接触，我们的初步数据表明，代谢信号可能会影响它们之间的适当通信。
Saccharomyces cerevisiae cells are killed by zymocin, a tRNase ribotoxin complex from Kluyveromyces lactis, which cleaves anticodons and inhibits protein synthesis. Zymocin’s action requires specific chemical modification of uridine bases in the anticodon wobble position (U34) by the Elongator complex (Elp1-Elp6). Hence, loss of anticodon modification in mutants lacking Elongator or related KTI (K. lactis Toxin Insensitive) genes protects against tRNA cleavage and confers resistance to the toxin. Here, we show that zymocin can be used as a tool to genetically analyse KTI12, a gene previously shown to code for an Elongator partner protein. From a kti12 mutant pool of zymocin survivors, we identify motifs in Kti12 that are functionally directly coupled to Elongator activity. In addition, shared requirement of U34 modifications for nonsense and missense tRNA suppression (SUP4; SOE1) strongly suggests that Kti12 and Elongator cooperate to assure proper tRNA functioning. We show that the Kti12 motifs are conserved in plant ortholog DRL1/ELO4 from Arabidopsis thaliana and seem to be involved in binding of cofactors (e.g., nucleotides, calmodulin). Elongator interaction defects triggered by mutations in these motifs correlate with phenotypes typical for loss of U34 modification. Thus, tRNA modification by Elongator appears to require physical contact with Kti12, and our preliminary data suggest that metabolic signals may affect proper communication between them.
登录
查看更多内容
影响因子：
14.8
作者：
Kelley LA;Mezulis S;Yates CM;Wass MN;Sternberg MJ
通讯作者：
Sternberg MJ
影响因子：
16.8
作者：
Glatt, Sebastian;Letoquart, Juliette;Mueller, Christoph W.
通讯作者：
Mueller, Christoph W.
影响因子：
5.7
作者：
Glatt, Sebastian;Zabel, Rene;Mueller, Christoph W.
通讯作者：
Mueller, Christoph W.
影响因子：
3.8
作者：
Jun SE;Cho KH;Hwang JY;Abdel-Fattah W;Hammermeister A;Schaffrath R;Bowman JL;Kim GT
通讯作者：
Kim GT
影响因子：
7.7
作者：
Dauden MI;Kosinski J;Kolaj-Robin O;Desfosses A;Ori A;Faux C;Hoffmann NA;Onuma OF;Breunig KD;Beck M;Sachse C;Séraphin B;Glatt S;Müller CW
通讯作者：
Müller CW