A base pair at the bottom of the anticodon stem is reciprocally preferred for discrimination of cognate tRNAs by Escherichia coli lysyl- and glutaminyl-tRNA synthetases.
A base pair at the bottom of the anticodon stem is reciprocally preferred for discrimination of cognate tRNAs by Escherichia coli lysyl- and glutaminyl-tRNA synthetases.
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DOI：
10.1093/nar/gkl414
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发表时间：
2006
影响因子：
14.9
通讯作者：
Yokogawa T
中科院分区：
文献类型：
--
作者：
Fukunaga J;Ohno S;Nishikawa K;Yokogawa T
虽然酵母琥珀抑制剂tRNATyr是一个很好的候选人的载体的非天然氨基酸进入蛋白质，轻微的misacylation与赖氨酸被发现发生在大肠杆菌蛋白质合成系统。虽然通过基因工程改造琥珀抑制基因tRNATyr的反密码子茎区可以抑制错糖基化，但发现对赖氨酸接受度最低的突变体tRNA反而接受痕量水平的谷氨酰胺。此外，谷氨酰胺接受各种tRNATyr转录物取代在反密码子茎区域的变化成反比的赖氨酸接受，类似于一个“跷跷板”。在该位点引入C31-G39碱基对对于降低赖氨酸接受和增加谷氨酰胺接受是最有效的。当将相同的取代引入大肠杆菌tRNALys转录物中时，赖氨酸接受活性降低了100倍，并且观察到微弱的谷氨酰胺接受。这些结果可能支持这样的想法，即在tRNA的反密码子茎中存在一些结构元件，这些结构元件不被在反密码子中具有相似识别位点的氨酰-tRNA合成酶共享，例如大肠杆菌赖氨酰-和氨酰-tRNA合成酶。
Although the yeast amber suppressor tRNATyr is a good candidate for a carrier of unnatural amino acids into proteins, slight misacylation with lysine was found to occur in an Escherichia coli protein synthesis system. Although it was possible to restrain the mislysylation by genetically engineering the anticodon stem region of the amber suppressor tRNATyr, the mutant tRNA showing the lowest acceptance of lysine was found to accept a trace level of glutamine instead. Moreover, the glutamine-acceptance of various tRNATyr transcripts substituted at the anticodon stem region varied in reverse proportion to the lysine-acceptance, similar to a ‘seesaw’. The introduction of a C31–G39 base pair at the site was most effective for decreasing the lysine-acceptance and increasing the glutamine-acceptance. When the same substitution was introduced into E.coli tRNALys transcripts, the lysine-accepting activity was decreased by 100-fold and faint acceptance of glutamine was observed. These results may support the idea that there are some structural element(s) in the anticodon stem of tRNA, which are not shared by aminoacyl-tRNA synthetases that have similar recognition sites in the anticodon, such as E.coli lysyl- and glutaminyl-tRNA synthetases.
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DOI：
10.1073/pnas.83.24.9373
发表时间：
1986-12-01
影响因子：
11.1
作者：
FREIER, SM;KIERZEK, R;TURNER, DH
通讯作者：
TURNER, DH
影响因子：
2.9
作者：
Guigou, L;Mirande, M
通讯作者：
Mirande, M
DOI：
10.1073/pnas.031488298
发表时间：
2001-02-27
影响因子：
11.1
作者：
Kowal, AK;Köhrer, C;RajBhandary, UL
通讯作者：
RajBhandary, UL
影响因子：
15
作者：
Wang, L;Magliery, TJ;Schultz, PG
通讯作者：
Schultz, PG
DOI：
10.1266/jjg.63.237
发表时间：
1988-06-01
期刊：
JAPANESE JOURNAL OF GENETICS
影响因子：
--
作者：
YAMAO, F;INOKUCHI, H;OZEKI, H
通讯作者：
OZEKI, H