PXD018793

PXD018793 is an original dataset announced via ProteomeXchange.

Title	Characterization of a novel two-component system TmrA/TmrB involved in multi-drug resistance of Pseudomonas fluorescens
Description	It has been widely reported that bacteria can sense environment changes and coordinates its behaviors via two-component systems （TCSs） to survive in the complicated niche. A TCS was mainly composed of a sensor kinase and a transcriptional factor, working as a signaling-sensor and a gene transcription regulator respectively. Thriving in diverse ecological niches, Pseudomonas fluorescens inherently resists a wide spectrum of antibiotics and the operon emhABC encoding an efflux pump plays a major role in the resistance to antibiotic and toxic compounds. Here, we identified that TmrA, a responsive regulator of the TmrA/TmrB two-component system, directly activated the expression of emhABC in P. fluorescens and a consensus motif recognized by TmrA was identified upstream of the emhABC promoter. Consistently, the tmrA deletion mutant (ΔtmrA) conferred significant decrease of resistance to multiple antibiotics. Moreover, deletion of intra-cellular kinase domain of TmrB resulted in decreased drug resistance similar to ΔTmrA mutant, inferring that phosphorylation of TmrA is essential for activation of emhABC expression. It was further demonstrated that in vitro phosphorylation of TmrA led to enhanced (>10 fold) binding affinity to the promoter region of emhABC. The knock-in TmrAD52A mutant, which bore the mutation of the conserved phosphorylation site across the TmrA homologues, showed partially decreased antibiotic resistance and in vitro phosphorylation of TmrAD52A protein also improved the binding to the emhABC promoter, suggesting there should be other unidentified phosphorylation sites yet essential to the regulation role of TmrA. Lastly, we performed the label-free quantification analysis of the whole proteome of the wild-type and ΔtmrA mutant strains and found that TmrA is likely to be a global regulator participating in multiple physiological processes including the multidrug resistance, secondary metabolite synthesis and nitrate assimilation. The characterization of other potential phosphorylation sites of TmrA is currently being undertaken.
HostingRepository	PRIDE
AnnounceDate	2021-10-08
AnnouncementXML	Submission_2021-10-08_02:04:08.707.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Diyin Li
SpeciesList	scientific name: Pseudomonas fluorescens F113; NCBI TaxID: 1114970;
ModificationList	monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2020-04-24 04:39:47	ID requested
⏵ 1	2021-10-08 02:04:09	announced

Dataset with its publication pending

submitter keyword: two-component system, TmrA/TmrB, efflux pumps, multi-antibiotic resistance, Pseudomonas fluorescens

Yongxing He
contact affiliation	Ministry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou 730000, P.R. China
contact email	heyx@lzu.edu.cn
lab head
Diyin Li
contact affiliation	Lanzhou University
contact email	lity16@lzu.edu.cn
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD018793
     1. Label: PRIDE project
     2. Name: Characterization of a novel two-component system TmrA/TmrB involved in multi-drug resistance of Pseudomonas fluorescens