摘要
东湖假单胞菌HYS对秀丽隐杆线虫具有较强毒性,但机制尚不完全了解。本研究旨在探究东湖假单胞菌HYS中σ因子SigW与反σ因子RsiW在其对秀丽隐杆线虫毒性中的作用。通过对东湖假单胞菌HYS减毒突变株突变位点序列进行生物信息分析,构建目的基因敲除突变株,借助酵母双杂交、荧光定量PCR以及假单胞菌-秀丽隐杆线虫互作等研究方法进行相关基因蛋白功能验证。结果发现:(1)基因rsiW与东湖假单胞菌HYS菌株对秀丽隐杆线虫毒性相关;(2)RsiW与基因组上邻近的SigW(σ24)是一对σ因子与反σ因子,在蛋白水平存在互作;(3)RsiW与SigW互作参与东湖假单胞菌HYS对秀丽隐杆线虫毒性作用且存在一定的剂量效应,即SigW表达相对较强时,存在减毒效应。该研究发现一对σ因子与反σ因子(SigW/RsiW)参与东湖假单胞菌HYS对秀丽隐杆线虫的毒性作用,为深入探讨假单胞菌毒力因子调控机制提供了新线索和思路。
Pseudomonas donghuensis HYS is toxic to Caenorhabditis elegans but its mechanism is not fully understood.This study aims to investigate the roles ofσfactor SigW and anti-σfactor RsiW in the toxicity of P.donghuensis HYS to C.elegans.Through the bioinformatics analysis of the mutation site sequence in the toxicity-reduced mutant of P.dongshanensis HYS,the target gene knockout mutant was constructed.Then with the help of research methods such as yeast two-hybrid,fluorescent quantitative PCR and P.dongshanensis-C.elegans interaction,etc.,the protein functions of related gene were verified.The results revealed that:(1)the gene rsiW was related to the toxicity of P.donghuensis HYS to C.elegans.(2)RsiW interacted with neighboring SigW(σ24)on the genome at the protein level,forming a pair ofσfactor and anti-σfactor.(3)The interaction between RsiW and SigW was involved in the toxicity of P.donghuensis HYS to C.elegans and there was a dose effect,i.e.,there was reducing effect when SigW expression was relatively strong.This study discovers that a pair ofσfactor and anti-σfactor are involved in the toxicity of P.donghuensis HYS to C.elegans and provides new clues and ideas for further exploring the mechanism of virulence factors of Pseudomonas.
作者
伍婷婷
桂哲
秦迎秋
谢志雄
WU Ting-ting;GUI Zhe;QIN Ying-qiu;XIE Zhi-xiong(College of Life Sciences,Wuhan University,Wuhan 430072)
出处
《生物技术通报》
CAS
CSCD
北大核心
2020年第10期142-149,共8页
Biotechnology Bulletin
基金
国家自然科学基金项目(31570090)
国家微生物资源平台项目(NIMR-2020-8)。