一种用于研究酿酒酵母中翻译重编码的体内双荧光素酶检测系统。
An in vivo dual-luciferase assay system for studying translational recoding in the yeast Saccharomyces cerevisiae.
作者信息
Harger Jason W, Dinman Jonathan D
机构信息
Department of Molecular Genetics, Microbiology and Immunology, UMDNJ Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
出版信息
RNA. 2003 Aug;9(8):1019-24. doi: 10.1261/rna.5930803.
Abstract
A new in vivo assay system has been developed to study programmed frameshifting in the yeast Saccharomyces cerevisiae. Frameshift signals are inserted between the Renilla and firefly luciferase reporter genes contained in a yeast expression vector and the two activities are directly measured from cell lysates in one tube. Similar to other bicistronic reporter systems, this one allows the efficient estimation of recoding efficiency by comparison of the normalized activity ratios from each luciferase protein. The assay system has been applied to HIV-1 and L-A directed programmed -1 frameshifting and Ty1 and Ty3 directed +1 frameshifting. The assay system is amenable to high-throughput screening.
摘要
已开发出一种新的体内检测系统，用于研究酿酒酵母中的程序性移码。移码信号插入酵母表达载体中包含的海肾荧光素酶和萤火虫荧光素酶报告基因之间，两种活性可直接在一管细胞裂解物中进行测量。与其他双顺反子报告系统类似，该系统通过比较每种荧光素酶蛋白的标准化活性比率，能够有效地评估重编码效率。该检测系统已应用于HIV-1和L-A介导的程序性-1移码以及Ty1和Ty3介导的+1移码。该检测系统适用于高通量筛选。
相似文献
1
An in vivo dual-luciferase assay system for studying translational recoding in the yeast Saccharomyces cerevisiae.一种用于研究酿酒酵母中翻译重编码的体内双荧光素酶检测系统。
RNA. 2003 Aug;9(8):1019-24. doi: 10.1261/rna.5930803.
2
A dual-luciferase reporter system for studying recoding signals.一种用于研究重编码信号的双荧光素酶报告系统。
RNA. 1998 Apr;4(4):479-86.
3
Dual luciferase assay system for rapid assessment of gene expression in Saccharomyces cerevisiae.用于快速评估酿酒酵母基因表达的双荧光素酶检测系统。
Eukaryot Cell. 2005 Sep;4(9):1539-49. doi: 10.1128/EC.4.9.1539-1549.2005.
4
Increased -1 ribosomal frameshifting efficiency by yeast prion-like phenotype [PSI+].酵母朊病毒样表型[PSI+]提高核糖体-1移码效率。
FEBS Lett. 2009 Feb 18;583(4):665-9. doi: 10.1016/j.febslet.2009.01.015. Epub 2009 Jan 21.
5
Decreased peptidyltransferase activity correlates with increased programmed -1 ribosomal frameshifting and viral maintenance defects in the yeast Saccharomyces cerevisiae.肽基转移酶活性降低与酿酒酵母中程序性-1核糖体移码增加及病毒维持缺陷相关。
RNA. 2003 Aug;9(8):982-92. doi: 10.1261/rna.2165803.
6
Expression of a MORN repeat protein from Euplotes octocarinatus requires a +1 programmed ribosomal frameshifting.来自八肋游仆虫的MORN重复蛋白的表达需要 +1程序性核糖体移码。
Biosci Biotechnol Biochem. 2017 Jul;81(7):1327-1334. doi: 10.1080/09168451.2017.1301804. Epub 2017 Mar 20.
7
The pokeweed antiviral protein specifically inhibits Ty1-directed +1 ribosomal frameshifting and retrotransposition in Saccharomyces cerevisiae.商陆抗病毒蛋白可特异性抑制酿酒酵母中Ty1介导的+1核糖体移码和逆转座作用。
J Virol. 1998 Feb;72(2):1036-42. doi: 10.1128/JVI.72.2.1036-1042.1998.
8
Systematic analysis of bicistronic reporter assay data.双顺反子报告基因检测数据的系统分析
Nucleic Acids Res. 2004 Nov 23;32(20):e160. doi: 10.1093/nar/gnh157.
9
Spermidine deficiency increases +1 ribosomal frameshifting efficiency and inhibits Ty1 retrotransposition in Saccharomyces cerevisiae.亚精胺缺乏会提高酿酒酵母中的 +1 核糖体移码效率并抑制 Ty1 逆转录转座。
Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):172-6. doi: 10.1073/pnas.91.1.172.
10
Evolutionary specialization of recoding: frameshifting in the expression of S. cerevisiae antizyme mRNA is via an atypical antizyme shift site but is still +1.重新编码的进化特化：酿酒酵母抗酶mRNA表达中的移码是通过一个非典型抗酶移码位点，但仍然是+1移码。
RNA. 2006 Mar;12(3):332-7. doi: 10.1261/rna.2245906. Epub 2006 Jan 23.
引用本文的文献
1
Comparative analyses of disease-linked missense mutations in the RNA exosome modeled in budding yeast reveal distinct functional consequences in translation.对在芽殖酵母中建模的RNA外泌体中与疾病相关的错义突变进行的比较分析揭示了翻译中不同的功能后果。
RNA. 2025 Jun 16;31(7):988-1012. doi: 10.1261/rna.080447.125.
2
Guidelines for minimal reporting requirements, design and interpretation of experiments involving the use of eukaryotic dual gene expression reporters (MINDR).涉及真核双基因表达报告基因实验的最低报告要求、设计与解释指南（MINDR）
Nat Struct Mol Biol. 2025 Mar;32(3):418-430. doi: 10.1038/s41594-025-01492-x. Epub 2025 Mar 3.
3
Phosphorylation of P-stalk proteins defines the ribosomal state for interaction with auxiliary protein factors.P柄蛋白的磷酸化决定了核糖体与辅助蛋白因子相互作用的状态。
EMBO Rep. 2024 Dec;25(12):5478-5506. doi: 10.1038/s44319-024-00297-1. Epub 2024 Oct 28.
4
Conserved 5-methyluridine tRNA modification modulates ribosome translocation.保守的 5-甲基尿嘧啶 tRNA 修饰调节核糖体易位。
Proc Natl Acad Sci U S A. 2024 Aug 27;121(35):e2401743121. doi: 10.1073/pnas.2401743121. Epub 2024 Aug 19.
5
Loss of Conserved rRNA Modifications in the Peptidyl Transferase Center Leads to Diminished Protein Synthesis and Cell Growth in Budding Yeast.核糖体肽酰转移酶中心保守的 rRNA 修饰缺失导致出芽酵母中蛋白质合成和细胞生长能力下降。
Int J Mol Sci. 2024 May 10;25(10):5194. doi: 10.3390/ijms25105194.
6
Shiftless Is a Novel Member of the Ribosome Stress Surveillance Machinery That Has Evolved to Play a Role in Innate Immunity and Cancer Surveillance.Shiftless 是核糖体应激监测机制中的一个新成员，它在进化过程中发挥了作用，参与了先天免疫和癌症监测。
Viruses. 2023 Nov 23;15(12):2296. doi: 10.3390/v15122296.
7
Comparative analyses of disease-linked missense mutations in the RNA exosome modeled in budding yeast reveal distinct functional consequences in translation.对芽殖酵母中建模的RNA外泌体中与疾病相关的错义突变进行的比较分析揭示了翻译中不同的功能后果。
bioRxiv. 2025 Mar 19:2023.10.18.562946. doi: 10.1101/2023.10.18.562946.
8
eEF2 diphthamide modification restrains spurious frameshifting to maintain translational fidelity.eEF2 二氢喋呤修饰抑制错义移码以维持翻译忠实性。
Nucleic Acids Res. 2023 Jul 21;51(13):6899-6913. doi: 10.1093/nar/gkad461.
9
A novel de novo variant in CASK causes a severe neurodevelopmental disorder that masks the phenotype of a novel de novo variant in EEF2.一种新的 CASK 基因中的从头变异导致了一种严重的神经发育障碍，掩盖了 EEF2 中一种新的从头变异的表型。
J Hum Genet. 2023 Aug;68(8):543-550. doi: 10.1038/s10038-023-01150-4. Epub 2023 Apr 18.
10
Interaction of the La-related protein Slf1 with colliding ribosomes maintains translation of oxidative-stress responsive mRNAs.La 相关蛋白 Slf1 与碰撞核糖体的相互作用维持氧化应激反应性 mRNAs 的翻译。
Nucleic Acids Res. 2023 Jun 23;51(11):5755-5773. doi: 10.1093/nar/gkad272.
本文引用的文献
1
Genetic Control of Biochemical Reactions in Neurospora.粗糙脉孢菌中生化反应的遗传控制。
Proc Natl Acad Sci U S A. 1941 Nov 15;27(11):499-506. doi: 10.1073/pnas.27.11.499.
2
An "integrated model" of programmed ribosomal frameshifting.程序化核糖体移码的“整合模型”
Trends Biochem Sci. 2002 Sep;27(9):448-54. doi: 10.1016/s0968-0004(02)02149-7.
3
New targets for antivirals: the ribosomal A-site and the factors that interact with it.抗病毒药物的新靶点：核糖体A位点及其相互作用因子。
Virology. 2002 Aug 15;300(1):60-70. doi: 10.1006/viro.2002.1567.
4
The frameshift signal of HIV-1 involves a potential intramolecular triplex RNA structure.HIV-1的移码信号涉及一种潜在的分子内三链RNA结构。
Proc Natl Acad Sci U S A. 2002 Apr 16;99(8):5331-6. doi: 10.1073/pnas.082102199.
5
Nonsense-mediated mRNA decay in Saccharomyces cerevisiae.酿酒酵母中的无义介导的mRNA降解
Gene. 2001 Aug 22;274(1-2):15-25. doi: 10.1016/s0378-1119(01)00552-2.
6
Ty1 retrotransposition and programmed +1 ribosomal frameshifting require the integrity of the protein synthetic translocation step.Ty1逆转座和程序性+1核糖体移码需要蛋白质合成易位步骤的完整性。
Virology. 2001 Jul 20;286(1):216-24. doi: 10.1006/viro.2001.0997.
7
The case against the involvement of the NMD proteins in programmed frameshifting.关于NMD蛋白参与程序性移码的反对观点。
RNA. 2000 Dec;6(12):1687-8. doi: 10.1017/s1355838200001874.
8
The case for the involvement of the Upf3p in programmed -1 ribosomal frameshifting.Upf3p参与程序性-1核糖体移码的情况。
RNA. 2000 Dec;6(12):1685-6. doi: 10.1017/s1355838200001886.
9
Nonsense-mediated decay mutants do not affect programmed -1 frameshifting.无义介导的衰变突变体不影响程序性-1移码。
RNA. 2000 Jul;6(7):952-61. doi: 10.1017/s1355838200000443.
10
A dual-luciferase reporter system for studying recoding signals.一种用于研究重编码信号的双荧光素酶报告系统。
RNA. 1998 Apr;4(4):479-86.
Zotero 插件