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叶绿体基于不同周转率的蛋白质的非 AUG 起始密码子，进化出了额外的翻译调控层。

Chloroplasts evolved an additional layer of translational regulation based on non-AUG start codons for proteins with different turnover rates.

机构信息

Plant Transformation Group, International Centre for Genetic Engineering and Biotechnology (ICGEB), New Delhi, 110067, India.

ICAR-Indian Institute of Maize Research, Delhi Unit, Pusa Campus, New Delhi, 110012, India.

出版信息

Sci Rep. 2023 Jan 17;13(1):896. doi: 10.1038/s41598-022-27347-9.

DOI:10.1038/s41598-022-27347-9
PMID:36650197
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9845219/
Abstract

Chloroplasts have evolved from photosynthetic cyanobacteria-like progenitors through endosymbiosis. The chloroplasts of present-day land plants have their own transcription and translation systems that show several similarities with prokaryotic organisms. A remarkable feature of the chloroplast translation system is the use of non-AUG start codons in the protein synthesis of certain genes that are evolutionarily conserved from Algae to angiosperms. However, the biological significance of such use of non-AUG codons is not fully understood. The present study was undertaken to unravel the significance of non-AUG start codons in vivo using the chloroplast genetic engineering approach. For this purpose, stable transplastomic tobacco plants expressing a reporter gene i.e. uidA (GUS) under four different start codons (AUG/UUG/GUG/CUG) were generated and β-glucuronidase (GUS) expression was compared. To investigate further the role of promoter sequences proximal to the start codon, uidA was expressed under two different chloroplast gene promoters psbA and psbC that use AUG and a non-AUG (GUG) start codons, respectively, and also showed significant differences in the DNA sequence surrounding the start codon. Further, to delineate the role of RNA editing that creates AUG start codon by editing non-AUG codons, if any, which is another important feature of the chloroplast transcription and translation system, transcripts were sequenced. In addition, a proteomic approach was used to identify the translation initiation site(s) of GUS and the N-terminal amino acid encoded when expressed under different non-AUG start codons. The results showed that chloroplasts use non-AUG start codons in combination with the translation initiation site as an additional layer of gene regulation to over-express proteins that are required at high levels due to their high rates of turnover.

摘要

叶绿体通过内共生从光合蓝细菌样祖先进化而来。现今陆地植物的叶绿体拥有自己的转录和翻译系统，这些系统与原核生物有一些相似之处。叶绿体翻译系统的一个显著特点是，在某些基因的蛋白质合成中使用非 AUG 起始密码子，这些基因从藻类到被子植物都是进化保守的。然而，这种非 AUG 密码子的使用的生物学意义尚未完全清楚。本研究采用叶绿体遗传工程方法，试图从体内阐明非 AUG 起始密码子的意义。为此，构建了四种不同起始密码子（AUG/UUG/GUG/CUG）下表达报告基因 uidA（GUS）的稳定转叶绿体烟草植物，并比较了 β-葡萄糖醛酸酶（GUS）的表达。为了进一步研究起始密码子附近启动子序列的作用，在两种不同的叶绿体基因启动子 psbA 和 psbC 下表达 uidA，它们分别使用 AUG 和非 AUG（GUG）起始密码子，并且在起始密码子周围的 DNA 序列上也存在显著差异。此外，为了阐明 RNA 编辑的作用，该编辑通过编辑非 AUG 密码子来创建 AUG 起始密码子，这是叶绿体转录和翻译系统的另一个重要特征，对转录本进行了测序。此外，还采用蛋白质组学方法来鉴定在不同非 AUG 起始密码子下表达时 GUS 的翻译起始位点和编码的 N 末端氨基酸。结果表明，叶绿体在结合翻译起始位点的情况下使用非 AUG 起始密码子，作为一种额外的基因调控层，以过表达由于其高周转率而需要高水平表达的蛋白质。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/a8eade6db099/41598_2022_27347_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/44a49e2d3ccb/41598_2022_27347_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/4839f20e8cca/41598_2022_27347_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/5ea8a951047a/41598_2022_27347_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/214d0ddfd2a7/41598_2022_27347_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/5038626e772e/41598_2022_27347_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/9c65b24ad6e7/41598_2022_27347_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/a8eade6db099/41598_2022_27347_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/44a49e2d3ccb/41598_2022_27347_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/4839f20e8cca/41598_2022_27347_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/5ea8a951047a/41598_2022_27347_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/214d0ddfd2a7/41598_2022_27347_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/5038626e772e/41598_2022_27347_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/9c65b24ad6e7/41598_2022_27347_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f666/9845219/a8eade6db099/41598_2022_27347_Fig7_HTML.jpg

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