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PXD030973

PXD030973 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleFRET monitoring of transcription factor activities in living bacteria
DescriptionAbstract: Bacteria adapt to the constantly changing environments largely by transcriptional regulation through the activities of various transcription factors (TFs). However, techniques that monitor the in situ TF-promoter interactions in living bacteria are lacking. Herein, we developed a whole-cell TF-promoter binding assay based on the intermolecular Förster resonance energy transfer (FRET) between a fluorescent unnatural amino acid CouA which is genetically encoded into defined sites in TFs and the live cell fluorescent nucleic acid stain SYTO 9. We show that this new FRET pair monitors the intricate TF-promoter interactions elicited by various types of signal transduction systems with specificity and sensitivity. Furthermore, the assay is applicable to identify novel modulators of the regulatory systems of interest and monitor TF activities in bacteria colonized in C. elegans. In conclusion, we established a tractable and sensitive TF-promoter binding assay in living bacteria which not only complements currently available approaches for DNA-protein interactions but also provides novel opportunities for functional annotation of bacterial signal transduction systems and studies of the bacteria-host interface
HostingRepositoryPRIDE
AnnounceDate2023-11-14
AnnouncementXMLSubmission_2023-11-14_07:36:36.582.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterPengchao Wang
SpeciesList scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Bacteria; NCBI TaxID: NCBITaxon:2;
ModificationListNo PTMs are included in the dataset
InstrumentOrbitrap Fusion Lumos
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-01-14 08:31:48ID requested
12022-10-13 05:34:47announced
22023-11-14 07:36:37announced2023-11-14: Updated project metadata.
Publication List
Wang P, Zhang G, Xu Z, Chen Z, Liu X, Wang C, Zheng C, Wang J, Zhang H, Yan A, Whole-cell FRET monitoring of transcription factor activities enables functional annotation of signal transduction systems in living bacteria. J Biol Chem, 298(8):102258(2022) [pubmed]
Keyword List
submitter keyword: Transcription factor
Promoter
FRET
Fluorescent Unnatural amino acid
Genetic codon expansion
Contact List
Dr. Aixin Yan
contact affiliationAssociate Professor, SBS, The University of Hong Kong
contact emailayan8@hku.hk
lab head
Pengchao Wang
contact affiliationThe Univerisity of Hong Kong
contact emailpengchao@connect.hku.hk
dataset submitter
Full Dataset Link List
Dataset FTP location
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