Simplification of the genetic code: restricted diversity of genetically encoded amino acids.
Simplification of the genetic code: restricted diversity of genetically encoded amino acids.
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DOI：
10.1093/nar/gks786
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发表时间：
2012-11-01
影响因子：
14.9
通讯作者：
Kiga D
中科院分区：
文献类型：
--
作者：
Kawahara-Kobayashi A;Masuda A;Araiso Y;Sakai Y;Kohda A;Uchiyama M;Asami S;Matsuda T;Ishitani R;Dohmae N;Yokoyama S;Kigawa T;Nureki O;Kiga D
在通用遗传密码进化的早期阶段，人们认为使用的氨基酸不到20个。尽管这一概念得到了广泛数据的支持，但与扩展密码的成功开发相比，具有有限规范氨基酸集的遗传密码的实际存在和功能尚未得到实验解决。在这里，我们构建了包含简化字母表的人工遗传密码。在其中一个密码子中，带有Trp反密码子的tRNAAla变体将丙氨酸重新分配给缺乏色氨酸的大肠杆菌S30无细胞翻译系统中未分配的UGG密码子。我们通过氨基酸组成分析、绿色荧光蛋白荧光测量和晶体结构测定，证实了这种缺乏Trp的密码合成蛋白质的效率和准确性与通用遗传密码相当。我们还证明了另一种编码，其中UGU/UGC密码子被分配给Ser，合成一种活性酶。这种方法不仅将提供对原始遗传密码的新见解，而且还将成为评估蛋白质进化早期阶段和改进药物的基本蛋白质工程工具。
At earlier stages in the evolution of the universal genetic code, fewer than 20 amino acids were considered to be used. Although this notion is supported by a wide range of data, the actual existence and function of the genetic codes with a limited set of canonical amino acids have not been addressed experimentally, in contrast to the successful development of the expanded codes. Here, we constructed artificial genetic codes involving a reduced alphabet. In one of the codes, a tRNAAla variant with the Trp anticodon reassigns alanine to an unassigned UGG codon in the Escherichia coli S30 cell-free translation system lacking tryptophan. We confirmed that the efficiency and accuracy of protein synthesis by this Trp-lacking code were comparable to those by the universal genetic code, by an amino acid composition analysis, green fluorescent protein fluorescence measurements and the crystal structure determination. We also showed that another code, in which UGU/UGC codons are assigned to Ser, synthesizes an active enzyme. This method will provide not only new insights into primordial genetic codes, but also an essential protein engineering tool for the assessment of the early stages of protein evolution and for the improvement of pharmaceuticals.
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DOI：
10.1126/science.1174343
发表时间：
2009-08-07
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Guo M;Chong YE;Beebe K;Shapiro R;Yang XL;Schimmel P
通讯作者：
Schimmel P
影响因子：
5.6
作者：
CRICK, FHC
通讯作者：
CRICK, FHC
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
DOI：
10.1023/b:jsfg.0000029204.57846.7d
发表时间：
2004-01-01
期刊：
Journal of Structural and Functional Genomics
影响因子：
--
作者：
Kigawa, Takanori;Yabuki, Takashi;Yokoyama, Shigeyuki
通讯作者：
Yokoyama, Shigeyuki
影响因子：
64.8
作者：
Ito, Takuhiro;Yokoyama, Shigeyuki
通讯作者：
Yokoyama, Shigeyuki