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.
RNA. 2003 Aug;9(8):1019-24. doi: 10.1261/rna.5930803.
2
A dual-luciferase reporter system for studying recoding signals.
RNA. 1998 Apr;4(4):479-86.
3
Dual luciferase assay system for rapid assessment of gene expression in Saccharomyces cerevisiae.
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+].
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.
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.
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.
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.
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.
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.
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.
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).
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
RNA. 2000 Dec;6(12):1685-6. doi: 10.1017/s1355838200001886.
9
Nonsense-mediated decay mutants do not affect programmed -1 frameshifting.
RNA. 2000 Jul;6(7):952-61. doi: 10.1017/s1355838200000443.
10
A dual-luciferase reporter system for studying recoding signals.
RNA. 1998 Apr;4(4):479-86.
Zotero 插件