PXD070275-1

PXD070275 is an original dataset announced via ProteomeXchange.

Title	Generation of an optimized Cumate circuit toolkit for tunable protein expression during in vitro and in vivo studies of Burkholderia cenocepacia: DIA experiment cumate vs Rhamnose
Description	Inducible gene expression systems are pivotal for dissecting bacterial physiology and virulence mechanisms. Across the Burkholderia genera, a limited range of inducible systems currently exist that show minimal impacts on the proteome and allow tight regulation. In this study, we engineer a set of cumate-inducible vectors for use in Burkholderia cenocepacia that offer minimal basal expression and the ability to control B. cenocepacia gene expression within Eukaryotic cells. Through mutagenesis-based studies of cumate circuits and the cumate regulator (CymR), we generate an optimized cumate circuit (PCymRC/CymRGV) which allows the tight and tunable control of protein expression within B. cenocepacia as assessed by fluorescent and protein O-linked glycosylation analysis. Using comparative proteomics, we demonstrate cumate induction leads to both reduced and orthogonal effects on B. cenocepacia compared to widely used rhamnose based induction systems. Leveraging the cell permeability of cumate and the generation of a CTX-based chromosomal integration vector, we show inducible control of protein expression is achievable during intracellular replication of B. cenocepacia. Finally, using the ability to control intracellular expression, we demonstrate the requirement of O-linked protein glycosylation for optimal B. cenocepacia intracellular replication. Combined, this work demonstrates that cumate inducible systems allows precise and tuneable gene expression in Burkholderia even within a host-pathogen context.
HostingRepository	PRIDE
AnnounceDate	2026-05-15
AnnouncementXML	Submission_2026-05-15_04:01:01.480.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Nichollas Scott
SpeciesList	scientific name: Burkholderia cenocepacia K56-2Valvano; NCBI TaxID: NEWT:985076;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2025-11-03 20:28:25	ID requested
⏵ 1	2026-05-15 04:01:01	announced

Dataset with its publication pending

submitter keyword: Burkholderia,Cumate

Nichollas E. Scott
contact affiliation	Department of Microbiology and Immunology, University of Melbourne, Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia
contact email	nichollas.scott@unimelb.edu.au
lab head
Nichollas Scott
contact affiliation	University of Melbourne
contact email	nichollas.scott@unimelb.edu.au
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD070275
     1. Label: PRIDE project
     2. Name: Generation of an optimized Cumate circuit toolkit for tunable protein expression during in vitro and in vivo studies of Burkholderia cenocepacia: DIA experiment cumate vs Rhamnose