title: Modulation of QscR, a quorum sensing receptor of Pseudomonas aeruginosa, by truncation of a signal binding domain.
abstract: In Pseudomonas aeruginosa, a multi-host pathogen, quorum sensing (QS) plays an essential role in pathogenesis, wherein LasR, QscR and RhlR, the QS regulators, control the expression of many virulence factors. In this study, we constructed a signal-binding-domain (SBD)-deleted QscR (QscR 160-237) to make a signal-independently-active form of QscR. However, QscR 160-237 that has only a DNA binding domain (DBD) was not fully active. It was able to bind to the target site in a signal-independent manner, but was not able to activate transcription of the target promoter. Since QscR 160-237 could interfere with binding of wild-type QscR (QscR wt) to its QscR binding site, we investigated the competition between QscR 160-237 and QscR wt on the QscR binding site in vivo and in vitro. When QscR wt and QscR 160-237 were independently co-expressed by two different inducers, increasing expression of QscR 160-237 interfered with QscR wt activity. This was verified by a competitive gel shift assay in vitro using purified QscR wt and QscR 160-237. Our results show that the SBD deletion makes QscR a partially active form that has only DNA binding ability, but it can interfere with QscR wt by competitive binding.
authors: Department of Pharmacy, College of Pharmacy, Laboratory of Microbiology, Pusan National University, Research Building 537, San 30, Jangjun-Dong, Geumjung-Gu, Busan 609-735, South Korea.
authors: Su-Jin
authors: SJ
authors: Park
authors: Hai-Bo
authors: HB
authors: Liu
authors: Sunghoon
authors: S
authors: Park
authors: Joon-Hee
authors: JH
authors: Lee
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D004252:DNA Mutational Analysis; D015964:Gene Expression Regulation, Bacterial; D011485:Protein Binding; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D053038:Quorum Sensing; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; C434259:QscR protein, Pseudomonas aeruginosa; D012097:Repressor Proteins