Deletion of the RluD pseudouridine synthase promotes SsrA peptide tagging of ribosomal protein S7.
Deletion of the RluD pseudouridine synthase promotes SsrA peptide tagging of ribosomal protein S7.
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DOI：
10.1111/j.1365-2958.2010.07467.x
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发表时间：
2011-01
影响因子：
3.6
通讯作者：
Hayes CS
中科院分区：
文献类型：
--
作者：
Schaub RE;Hayes CS
RluD催化50 S核糖体亚基的螺旋69内的三个假尿苷残基的形成。阻碍69与30 S亚基上的解码中心进行重要接触，并且最近显示rluD的缺失干扰大肠杆菌中的翻译终止。在这里，我们表明，删除rluD增加tmRNA活性的核糖体经历释放因子2（RF 2）介导的终止在UGA终止密码子。引人注目的是，tRNA介导的两种蛋白质（核糖体蛋白S7和LacI）的SsrA肽标记在ΔrluD细胞中显著增加。S7标记是由于在E. coli K-12菌株。将编码S7的rpsG基因从E. coli B菌株在ΔrluD背景下取消了S7标记，并部分补充了突变体的缓慢生长表型。此外，K-12 prf B基因（编码RF 2）与B菌株等位基因的交换大大减少了ΔrluD细胞中的标记。与E. coliK-12细胞中rluD的缺失。coli B菌株无生长表型。这些发现表明，最初观察到的rluD表型是由与大肠杆菌中发现的rpsG和prfB等位基因的合成相互作用引起的。coli K-12菌株。
RluD catalyzes formation of three pseudouridine residues within helix 69 of the 50S ribosome subunit. Helix 69 makes important contacts with the decoding center on the 30S subunit and deletion of rluD was recently shown to interfere with translation termination in Escherichia coli. Here, we show that deletion of rluD increases tmRNA activity on ribosomes undergoing release factor 2 (RF2)-mediated termination at UGA stop codons. Strikingly, tmRNA-mediated SsrA peptide tagging of two proteins, ribosomal protein S7 and LacI, was dramatically increased in ΔrluD cells. S7 tagging was due to a unique C-terminal peptide extension found in E. coli K-12 strains. Introduction of the rpsG gene (encoding S7) from an E. coli B strain abrogated S7 tagging in the ΔrluD background, and partially complemented the mutant’s slow-growth phenotype. Additionally, exchange of the K-12 prfB gene (encoding RF2) with the B strain allele greatly reduced tagging in ΔrluD cells. In contrast to E. coli K-12 cells, deletion of rluD in an E. coli B strain resulted in no growth phenotype. These findings indicate that the originally observed rluD phenotypes result from synthetic interactions with rpsG and prfB alleles found within E. coli K-12 strains.
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影响因子：
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作者：
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