Premature termination codon: a tunable protein translation approach

Cellular protein–protein interactions are largely dependent on the activities of signaling proteins. Here, we present a technique to tune gene expression at translation level based on G418-inducible readthrough premature termination codon (PTC-on). To demonstrate how this PTC-on can control the expr...

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Main Authors: Xiyao Cheng, Ting Zhou, Zixin Yang, Jingjing Zhou, Meng Gao, Yongqi Huang, Zhengding Su
Format: Article
Language:English
Published: Taylor & Francis Group 2022-08-01
Series:BioTechniques
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Online Access:https://www.future-science.com/doi/10.2144/btn-2022-0046
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author Xiyao Cheng
Ting Zhou
Zixin Yang
Jingjing Zhou
Meng Gao
Yongqi Huang
Zhengding Su
author_facet Xiyao Cheng
Ting Zhou
Zixin Yang
Jingjing Zhou
Meng Gao
Yongqi Huang
Zhengding Su
author_sort Xiyao Cheng
collection DOAJ
description Cellular protein–protein interactions are largely dependent on the activities of signaling proteins. Here, we present a technique to tune gene expression at translation level based on G418-inducible readthrough premature termination codon (PTC-on). To demonstrate how this PTC-on can control the expression level of a cellular signaling protein to regulate signal transduction, we settled a p53 PTC-on system in p53-null H1299 cells. After treating with G418, the cells expressed full-length p53 protein in a dose-dependent manner. We further demonstrated to use this PTC-on approach to dissect p53-dependent and p53-independent apoptosis in response to the DNA double strand breaks in H1299 cells. In principle, the PTC-on can be used as a general approach for exploring the functions of any other signaling proteins.
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1940-9818
language English
publishDate 2022-08-01
publisher Taylor & Francis Group
record_format Article
series BioTechniques
spelling doaj-art-829a8aae5f3b47078ff904973b24c79e2025-08-20T02:25:34ZengTaylor & Francis GroupBioTechniques0736-62051940-98182022-08-01732808910.2144/btn-2022-0046Premature termination codon: a tunable protein translation approachXiyao Cheng0Ting Zhou1Zixin Yang2Jingjing Zhou3Meng Gao4Yongqi Huang5Zhengding Su61School of Light Industry and Food Engineering, Guanxi University, No. 100, Daxuedong Road, Xixiangtang District, Nanning, Guangxi, 530004, China2Protein Engineering & Biopharmaceutical Sciences Laboratory, Hubei University of Technology, 28 Nanli Road, Wuhan, Hubei, 430068, China2Protein Engineering & Biopharmaceutical Sciences Laboratory, Hubei University of Technology, 28 Nanli Road, Wuhan, Hubei, 430068, China2Protein Engineering & Biopharmaceutical Sciences Laboratory, Hubei University of Technology, 28 Nanli Road, Wuhan, Hubei, 430068, China2Protein Engineering & Biopharmaceutical Sciences Laboratory, Hubei University of Technology, 28 Nanli Road, Wuhan, Hubei, 430068, China2Protein Engineering & Biopharmaceutical Sciences Laboratory, Hubei University of Technology, 28 Nanli Road, Wuhan, Hubei, 430068, China2Protein Engineering & Biopharmaceutical Sciences Laboratory, Hubei University of Technology, 28 Nanli Road, Wuhan, Hubei, 430068, ChinaCellular protein–protein interactions are largely dependent on the activities of signaling proteins. Here, we present a technique to tune gene expression at translation level based on G418-inducible readthrough premature termination codon (PTC-on). To demonstrate how this PTC-on can control the expression level of a cellular signaling protein to regulate signal transduction, we settled a p53 PTC-on system in p53-null H1299 cells. After treating with G418, the cells expressed full-length p53 protein in a dose-dependent manner. We further demonstrated to use this PTC-on approach to dissect p53-dependent and p53-independent apoptosis in response to the DNA double strand breaks in H1299 cells. In principle, the PTC-on can be used as a general approach for exploring the functions of any other signaling proteins.https://www.future-science.com/doi/10.2144/btn-2022-0046apoptosiscell cycle arrestgene expressionMMSp53premature termination codon
spellingShingle Xiyao Cheng
Ting Zhou
Zixin Yang
Jingjing Zhou
Meng Gao
Yongqi Huang
Zhengding Su
Premature termination codon: a tunable protein translation approach
BioTechniques
apoptosis
cell cycle arrest
gene expression
MMS
p53
premature termination codon
title Premature termination codon: a tunable protein translation approach
title_full Premature termination codon: a tunable protein translation approach
title_fullStr Premature termination codon: a tunable protein translation approach
title_full_unstemmed Premature termination codon: a tunable protein translation approach
title_short Premature termination codon: a tunable protein translation approach
title_sort premature termination codon a tunable protein translation approach
topic apoptosis
cell cycle arrest
gene expression
MMS
p53
premature termination codon
url https://www.future-science.com/doi/10.2144/btn-2022-0046
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AT tingzhou prematureterminationcodonatunableproteintranslationapproach
AT zixinyang prematureterminationcodonatunableproteintranslationapproach
AT jingjingzhou prematureterminationcodonatunableproteintranslationapproach
AT menggao prematureterminationcodonatunableproteintranslationapproach
AT yongqihuang prematureterminationcodonatunableproteintranslationapproach
AT zhengdingsu prematureterminationcodonatunableproteintranslationapproach