Laboratory of Molecular Genetics, Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba, Japan.
FEBS J. 2023 Jul;290(13):3480-3489. doi: 10.1111/febs.16760. Epub 2023 Mar 6.
The CGA codon is a rare codon in Saccharomyces cerevisiae and is known to be inefficiently decoded by wobble pairing with Arg-tRNA(ICG). The tRNA (ICG) is post-transcriptionally edited from tRNA (ACG) by the anticodon first adenosine deamination enzyme Tad2/Tad3 complex. Experimental consecutive CGA codons cause ribosome stalling to result in the reduction of the encoding protein product. In this study, the additional supply of tRNA (ACG) genes that produce decoding Arg-tRNA(ICG) promoted the product level from the CGA12-luc reporter, revealing that the product reduction is essentially due to inefficient decoding and deficiency in the tRNA supply. The mature tRNA (ICG) and the precursor tRNA (ACG) ratios examined for cellular tRNA fraction revealed that the tRNA (ICG) ratio is maintained at less than 30% and is responsive to the Tad2/Tad3 expression level.
CGA 密码子在酿酒酵母中是一种罕见的密码子,已知其与 Arg-tRNA(ICG)的摆动配对解码效率较低。tRNA(ICG)是通过反密码子第一个腺苷脱氨酶 Tad2/Tad3 复合物从 tRNA(ACG)转录后编辑而来的。实验中连续的 CGA 密码子导致核糖体停滞,从而减少编码蛋白产物。在这项研究中,额外供应产生解码 Arg-tRNA(ICG)的 tRNA(ACG)基因促进了 CGA12-luc 报告基因的产物水平,表明产物减少主要是由于解码效率低下和 tRNA 供应不足。对细胞 tRNA 分数进行的成熟 tRNA(ICG)和前体 tRNA(ACG)比例检查表明,tRNA(ICG)比例保持在 30%以下,并对 Tad2/Tad3 表达水平有响应。