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植物来源的半胱氨酸tRNA作为新型UGA抑制因子。

Cysteine tRNAs of plant origin as novel UGA suppressors.

作者信息

Urban C, Beier H

机构信息

Institut für Biochemie, Bayerische Julius-Maximilians-Universität, Würzburg, Germany.

出版信息

Nucleic Acids Res. 1995 Nov 25;23(22):4591-7. doi: 10.1093/nar/23.22.4591.

DOI:10.1093/nar/23.22.4591
PMID:8524647
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC307430/
Abstract

We have isolated and sequenced chloroplast (chl) and cytoplasmic (cyt) cysteine tRNAs from Nicotiana rustica. Both tRNAs carry a GCA anticodon but beyond that differ considerably in their nucleotide sequences. One obvious distinction resides in the presence of N6-isopentenyladenosine (i6A) and 1-methylguanosine (m1G) at position 37 in chl and cyt tRNA(Cys) respectively. In order to study the potential suppressor activity of tRNAs(Cys) we used in vitro synthesized zein mRNA transcripts in which an internal UGA stop codon had been placed in either the tobacco rattle virus (TRV)- or tobacco mosaic virus (TMV)-specific codon context. In vitro translation was carried out in a messenger- and tRNA-dependent wheat germ extract. Both tRNA(Cys) isoacceptors stimulate read-through over the UGA stop codon, however, chl tRNA(GCA)Cys is more efficient than the cytoplasmic counterpart. The UGA in the two viral codon contexts is suppressed to about the same extent by either of the two tRNAs(Cys), whereas UGA in the beta-globin context is not recognized at all. The interaction of tRNA(GCA)Cys with UGA requires an unconventional G:A base pair in the wobble position, as postulated earlier for plant tRNA(G psi A)Tyr misreading the UAA stop codon. This is the first case that a cysteine-accepting tRNA has been characterized as a natural UGA suppressor.

摘要

我们从黄花烟草中分离并测序了叶绿体（chl）和细胞质（cyt）中的半胱氨酸tRNA。两种tRNA都携带GCA反密码子，但除此之外，它们的核苷酸序列有很大差异。一个明显的区别在于，chl tRNA（Cys）和cyt tRNA（Cys）的第37位分别存在N6-异戊烯基腺苷（i6A）和1-甲基鸟苷（m1G）。为了研究tRNA（Cys）的潜在抑制活性，我们使用了体外合成的玉米醇溶蛋白mRNA转录本，其中内部UGA终止密码子被置于烟草脆裂病毒（TRV）或烟草花叶病毒（TMV）特异性密码子背景中。体外翻译在依赖信使RNA和tRNA的小麦胚提取物中进行。两种tRNA（Cys）同工受体都能刺激UGA终止密码子的通读，然而，chl tRNA（GCA）Cys比细胞质中的对应物更有效。两种tRNA（Cys）对两种病毒密码子背景中的UGA抑制程度大致相同，而β-珠蛋白背景中的UGA根本不被识别。tRNA（GCA）Cys与UGA的相互作用在摆动位置需要一个非常规的G:A碱基对，正如之前假设的植物tRNA（GψA）Tyr误读UAA终止密码子的情况。这是首次将一种接受半胱氨酸的tRNA鉴定为天然UGA抑制子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cafc/307430/69f4caf8cfdc/nar00022-0071-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cafc/307430/c6af2abb3c5e/nar00022-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cafc/307430/c3246f278430/nar00022-0071-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cafc/307430/69f4caf8cfdc/nar00022-0071-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cafc/307430/c6af2abb3c5e/nar00022-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cafc/307430/c3246f278430/nar00022-0071-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cafc/307430/69f4caf8cfdc/nar00022-0071-b.jpg

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本文引用的文献

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