σFactor SigW/Anti-σFactor RsiW参与东湖假单胞菌HYS对秀丽隐杆线虫毒性作用

σFactor SigW/Anti-σFactor Involving in the Toxicity of Pseudomonas donghuensis HYS to Caenorhabditis elegans

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.