RNA-seq for MvfR, RhlR and QscR

Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen of humans, most notably those with cystic fibrosis or whose immune systems are compromised. As a global regulator, MvfR,RhlR and QscR have been characterized to control a group of virulence-related factors and quorum sensing (QS) genes in P. aeruginosa, but its detailed molecular regulatory mechanism is largely elusive. To further gain insights into the direct targets of MvfR,RhlR and QscR in viv, this study focused on identifying the potential targets of MvfR,RhlR and QscR directly regulating QS system and other vireulence pathways. We performed a chromatin immunoprecipitation coupled to high-throughput sequencing (ChIP-seq) assay that identified 221, 30 and 1 binding sites of MvfR,RhlR and QscR. The direct regulation of these genes by MvfR,RhlR and QscR have been biochemically and genetically verified. The results indicated that MvfR,RhlR and QscR had direct and profound effects on QS, which provides new cues to better understand the detailed regulatory networks of virulence. RNA-seq for MvfR,RhlR and QscR

铜绿假单胞菌（Pseudomonas aeruginosa）是一种人类革兰氏阴性机会致病菌，尤其易感染囊性纤维化患者或免疫系统受损人群。作为一类全局调控因子，MvfR、RhlR与QscR已被证实可调控铜绿假单胞菌中的一系列毒力相关因子与群体感应（quorum sensing, QS）基因，但其具体的分子调控机制仍尚未完全阐明。为进一步解析MvfR、RhlR与QscR在体内的直接靶标，本研究聚焦于筛选可直接调控QS系统及其他毒力通路的MvfR、RhlR与QscR潜在靶标。本研究通过染色质免疫共沉淀结合高通量测序（ChIP-seq）实验，分别鉴定得到MvfR、RhlR与QscR的221、30及1个结合位点。上述基因受MvfR、RhlR与QscR的直接调控作用已通过生化与遗传学实验得到验证。研究结果显示，MvfR、RhlR与QscR对QS系统具有直接且显著的调控作用，这为进一步阐明毒力相关调控网络提供了新的线索。本研究同时开展了针对MvfR、RhlR与QscR的RNA测序（RNA-seq）实验。