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原核生物起始 tRNA(fMet)在核糖体 P 位选择的准确性中，起始 tRNA 基因多拷贝的关键作用。

Crucial contribution of the multiple copies of the initiator tRNA genes in the fidelity of tRNA(fMet) selection on the ribosomal P-site in Escherichia coli.

机构信息

Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560012, India.

出版信息

Nucleic Acids Res. 2011 Jan;39(1):202-12. doi: 10.1093/nar/gkq760. Epub 2010 Aug 26.

DOI:10.1093/nar/gkq760
PMID:20798174
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3017606/
Abstract

The accuracy of the initiator tRNA (tRNA(fMet)) selection in the ribosomal P-site is central to the fidelity of protein synthesis. A highly conserved occurrence of three consecutive G-C base pairs in the anticodon stem of tRNA(fMet) contributes to its preferential selection in the P-site. In a genetic screen, using a plasmid borne copy of an inactive tRNA(fMet) mutant wherein the three G-C base pairs were changed, we isolated Escherichia coli strains that allow efficient initiation with the tRNA(fMet) mutant. Here, extensive characterization of two such strains revealed novel mutations in the metZWV promoter severely compromising tRNA(fMet) levels. Low cellular abundance of the chromosomally encoded tRNA(fMet) allows efficient initiation with the tRNA(fMet) mutant and an elongator tRNA(Gln), revealing that a high abundance of the cellular tRNA(fMet) is crucial for the fidelity of initiator tRNA selection on the ribosomal P-site in E. coli. We discuss possible implications of the changes in the cellular tRNA(fMet) abundance in proteome remodeling.

摘要

起始 tRNA（tRNA(fMet)) 在核糖体 P 位的准确性对蛋白质合成的忠实性至关重要。tRNA(fMet) 的反密码子茎中三个连续的 G-C 碱基对的高度保守存在有助于其在 P 位的优先选择。在使用质粒携带的失活 tRNA(fMet) 突变体的遗传筛选中，我们分离出了允许该 tRNA(fMet) 突变体有效起始的大肠杆菌菌株。在这里，对两个这样的菌株进行了广泛的表征，发现 metZWV 启动子中的新型突变严重影响了 tRNA(fMet) 的水平。染色体编码的 tRNA(fMet) 的细胞丰度低允许与 tRNA(fMet) 突变体和延伸 tRNA(Gln) 有效起始，这表明细胞 tRNA(fMet) 的高丰度对于大肠杆菌核糖体 P 位起始 tRNA 选择的忠实性至关重要。我们讨论了细胞内 tRNA(fMet) 丰度变化在蛋白质组重塑中的可能影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/ffcb4712922b/gkq760f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/80291037b7f9/gkq760f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/e37245be2bdc/gkq760f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/ae47859579e9/gkq760f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/7b44ecb26c85/gkq760f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/89fbeeb4b570/gkq760f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/ffcb4712922b/gkq760f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/80291037b7f9/gkq760f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/e37245be2bdc/gkq760f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/ae47859579e9/gkq760f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/7b44ecb26c85/gkq760f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/89fbeeb4b570/gkq760f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/851b/3017606/ffcb4712922b/gkq760f6.jpg

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