阿维链霉菌中ECF-Sig25的调控通路  

Regulatory Pathway of ECF-Sig25 in Streptomyces avermitilis

作　　者：罗帅 王晓杰[1] 张虎成[1] 刘霖颖 简伟杰 朱华[2] 朱建亚 LUO Shuai;WANG Xiaojie;ZHANG Hucheng;LIU Linying;JIAN Weijie;ZHU Hua;ZHU Jianya(College of Biological Engineering,Beijing Polytechnic,Beijing 100176,China;Beijing Fisheries Research Institute,Beijing 100068,China)

机构地区：[1]北京电子科技职业学院生物工程学院,北京100176 [2]北京市水产科学研究所,北京100068

出　　处：《中国农业科技导报》2024年第6期122-132,共11页Journal of Agricultural Science and Technology

基　　金：北京电子科技职业学院科技类重点课题项目(2024X016-KXZ,2022X011-KXZ)。

摘　　要：ECF-Sigma因子和双组份调控系统连锁调控在细菌适应不同的生存环境和压力条件中起着重要作用。为研究阿维链霉菌中的ECF-sig25基因,通过孢子点接试验检测Sig25对胁迫的响应,通过平板划线分析其对形态分化的影响,通过生物信息学、RT-qPCR等验证Sig25的靶基因,以探索Sig25的调控通路。结果显示,ECF-sig25基因上游存在1个双组份调控系统基因smrAB;且上游调控元件SmrA的磷酸化严格依赖于SmrB。压力胁迫试验显示,Sig25响应碱、渗透压、去污剂和热激胁迫,不响应氧化、酸和EDTA胁迫。生物信息学预测、RT-qPCR和EMSA(electrophoretic mobility shift assay)揭示,Sig25直接调控whiG、SAV3434、SAV4621、SAV5010、SAV6537、SAV7165、gdhA1、glnA2和amtB1基因的转录,这些基因分别参与形态分化、甲基供给、DNA复制、信号传导、次级代谢和氮代谢等生理过程。平板划线结果显示,缺失sig25不影响阿维链霉菌的形态分化。The linkage regulation of extracytoplasmic function(ECF)sigma factors and two-component system(TCS)play essential roles for bacteria adaptation to various environment and stress condition.In order to study the ECFsig25 gene in Streptomyces avermitilis and explore the regulatory pathway of Sig25,the response of Sig25 to stress was detected by spore spot test,its influence on morphological differentiation was analyzed by plate culture,and the target gene of Sig25 was verified by bioinformatics and RT-qPCR.The results showed that TCS gene smrAB was located upstream of ECF-sig25,and the phosphorylation of SmrA depended solely on SmrB.Stress tests showed that Sig25 responded to high pH,osmotic stress,SDS and heat stress,but not to oxidative stress,acid stress and EDTA stress.Bioinformatic prediction,RT-qPCR and electrophoretic mobility shift assay(EMSA)results revealed that Sig25 directly regulated transcription of whiG,SAV3434,SAV4621,SAV5010,SAV6537,gdhA1,glnA2 and amtB1,which were involved in morphological development,methyl group supply,DNA replication,signal transduction,secondary metabolism and nitrogen metabolism.The plate culture showed that deletion of sig25 had no effect on morphological developmentin of S.avermitilis.

关 键 词：阿维链霉菌 ECF-Sigma因子 Sig25 双组份调控系统 SmrAB 

分 类 号：Q78[生物学—分子生物学]