Functional importance of Ψ38 and Ψ39 in distinct tRNAs, amplified for tRNAGln(UUG) by unexpected temperature sensitivity of the s2U modification in yeast.
Functional importance of Ψ38 and Ψ39 in distinct tRNAs, amplified for tRNAGln(UUG) by unexpected temperature sensitivity of the s2U modification in yeast.
复制标题
DOI：
10.1261/rna.048173.114
复制
发表时间：
2015-02
期刊：
影响因子：
--
通讯作者：
Phizicky EM
中科院分区：
文献类型：
--
作者：
Han L;Kon Y;Phizicky EM
tRNA的大量修饰在控制tRNA结构和翻译中起着核心作用。反密码子环内部和周围的修饰通常在mRNA解码和维持其阅读框中起关键作用。反密码子茎环中的残基U38和U39经常被广泛保守的伪尿嘧啶酶TruA/Pus3家族成员修饰为伪尿嘧啶（Ψ）。我们研究了酵母pus3Δ突变体温度敏感性的原因，发现尽管在至少19种表征的细胞质tRNA物种上发现Ψ38或Ψ39，但温度敏感性主要是由于tRNAGln（UUG）功能不佳，通常具有Ψ38。进一步研究发现，在高温下，tRNAGln（UUG）和另外两个细胞质种的mcm5s2U34的s2U片段水平显著降低，在亲本菌株BY4741和广泛使用的菌株W303中都出现了s2U水平的降低，而BY4741在低至33℃的温度下也能检测到s2U片段的降低。对Ψ38,39的作用的进一步研究提供了Ψ38在允许温度下对tRNAGln（UUG）的功能很重要的证据，并表明Ψ39对trm10Δ pus3Δ突变体中的tRNATrp（CCA）和tRNALeu（CAA）作为UAG无意义抑制因子的功能很重要。这些结果为Ψ38和Ψ39在特定trna中的重要作用提供了证据，并确定了在相对温和的生长条件下，摆动位置的修改会发生变化。
The numerous modifications of tRNA play central roles in controlling tRNA structure and translation. Modifications in and around the anticodon loop often have critical roles in decoding mRNA and in maintaining its reading frame. Residues U38 and U39 in the anticodon stem–loop are frequently modified to pseudouridine (Ψ) by members of the widely conserved TruA/Pus3 family of pseudouridylases. We investigate here the cause of the temperature sensitivity of pus3Δ mutants of the yeast Saccharomyces cerevisiae and find that, although Ψ38 or Ψ39 is found on at least 19 characterized cytoplasmic tRNA species, the temperature sensitivity is primarily due to poor function of tRNAGln(UUG), which normally has Ψ38. Further investigation reveals that at elevated temperatures there are substantially reduced levels of the s2U moiety of mcm5s2U34 of tRNAGln(UUG) and the other two cytoplasmic species with mcm5s2U34, that the reduced s2U levels occur in the parent strain BY4741 and in the widely used strain W303, and that reduced levels of the s2U moiety are detectable in BY4741 at temperatures as low as 33°C. Additional examination of the role of Ψ38,39 provides evidence that Ψ38 is important for function of tRNAGln(UUG) at permissive temperature, and indicates that Ψ39 is important for the function of tRNATrp(CCA) in trm10Δ pus3Δ mutants and of tRNALeu(CAA) as a UAG nonsense suppressor. These results provide evidence for important roles of both Ψ38 and Ψ39 in specific tRNAs, and establish that modification of the wobble position is subject to change under relatively mild growth conditions.
登录
查看更多内容
影响因子：
56.9
作者：
Gerber, AP;Keller, W
通讯作者：
Keller, W
影响因子：
5.6
作者：
Auffinger, P;Westhof, E
通讯作者：
Westhof, E
影响因子：
4.5
作者：
Gustavsson, Marie;Ronne, Hans
通讯作者：
Ronne, Hans
影响因子：
4.5
作者：
Dean, Kimberly M.;Grayhack, Elizabeth J.
通讯作者：
Grayhack, Elizabeth J.
影响因子：
14.9
作者：
Davis, DR
通讯作者：
Davis, DR