文献类型：期刊文献

中文题名：DNA硫修饰结合蛋白识别序列的表征与拓展促进新型核酸检测技术发展

英文题名：Profile and relaxation of sequence-specificity of DNA sulfur binding domains facilitate new nucleic acid detection platform

作者：帅玉婷[1];许安安[1];李甲乙[2];韩照晰[1];马蒂尼[1,2];段海容[1];王馨叶[1];蒋岚[1];张敬宇[1];谭高翼[1];刘雪婷[1];赵一雷[2];童垚俊[2];王申林[1];贺新义[2];邓子新[2];刘光[1,2];张立新[1]

机构：[1]East China Univ Sci & Technol ECUST, Sch Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, State Key Lab Microbial Metab, Joint Int Res Lab Metab & Dev Sci, Shanghai 200240, Peoples R China

年份：2023

卷号：68

期号：16

起止页码：1752

中文期刊名：Science Bulletin

外文期刊名：科学通报（英文版）

收录：CSTPCD;;EI(收录号：20233014447259);Scopus;WOS:【SCI-EXPANDED(收录号:WOS:001066278700001)】；CSCD:【CSCD2023_2024】；PubMed;

基金：This work was supported by the National Key Research and Development Program of China (2020YFA0907800, 2022YFC3400200, and 2022YFA0912200) , the National Natural Science Foundation of China (31900060) , the Shanghai Pilot Program for Basic Research-Shanghai Jiao Tong University (21TQ1400204) , and the Natural Science Foundation of Shanghai (20ZR1414500) .

语种：中文

中文关键词：核酸检测技术;限制性核酸内切酶;SBD;序列模式;识别序列;防御系统;硫原子;抗氧化应激;

摘要：DNA硫修饰是一种DNA磷酸骨架上的非桥接氧原子被硫原子所取代的修饰,属于原核生物表观遗传标志物之一,它对细菌防御系统、抗氧化应激和基因调控等生命过程具有重要影响.DNA硫修饰可以被硫修饰依赖限制性核酸内切酶的硫结合域SBD特异性识别,这使其具有被开发为新型生物技术工具的潜力.然而目前关于SBD蛋白识别序列的研究仍比较粗浅且仅限于少数天然序列模式,这极大限制了其应用开发.因此,我们建立了一种针对SBD蛋白识别序列表征的技术(SBS-seq),用于对SBD的完整识别序列进行高分辨率表征.而且,采用在DNA上串联多个硫修饰的方案来提高SBD的结合亲和力并拓展了其序列识别范围.在此基础上,我们进一步优化了基于SBD和硫修饰DNA的新型核酸检测技术.本工作为硫修饰依赖限制性核酸内切酶的研究提供了思路和范例,并促进了硫修饰DNA相关生物技术的应用.
DNA phosphorothioate(PT)-modification,with a non-bridging oxygen in the phosphodiester backbone substituted by sulfur,is an epigenetic marker in prokaryotes and involved in the bacterial defense system,anti-oxidative stress,and gene regulation[1–4].PT-modification could be specifically recognized by sulfur binding domains(SBD)of PT-dependent restriction endonucleases(REases)[5,6],making it a potential tool for enabling biotechnology development[7].However,the unclear recognition sequence-range of SBDs limits the application.Here,we report a technique named sulfur binding specificity-sequencing(SBS-seq)for highresolution characterization of the complete sequence-range of SBDs.We then employed tandem PT-modifications to improve the binding affinity and extend the sequence-range.The strong affinity and broad range facilitate a new nucleic acid detection platform based on SBD and PT-DNA.Our work provides insights for research on modification-dependent REases and facilitates biotechnology applications of PT-DNA.