PXD063323

PXD063323 is an original dataset announced via ProteomeXchange.

Title	Bacillus velezensis LoaP Promotes Antitermination of Antibiotic Synthesis Operons by Antagonizing NusA
Description	The transcription factor NusG is essential for the bacterial transcription elongation complex (TEC). During elongation, NusG dynamically interacts with other proteins, affecting transcription-translation coupling, processivity, and termination. NusG is integral to antitermination complexes, where it associates with other factors to modify the TEC such that it bypasses termination sites. In Gammaproteobacteria, these complexes enhance the expression of ribosomal RNA operons, bacteriophage operons, and CRISPR arrays. Specialized NusG paralogs, such as LoaP, RfaH, and UpxY, are believed to form their own unique antitermination complexes. RfaH, the only well-studied paralog, binds to a paused TEC via a specific nontemplate DNA sequence. The RfaH-bound TEC then enhances transcription-translation coupling while also preventing Rho termination. However, it is unclear whether the other NusG paralogs use similar mechanisms, nor have antitermination complexes been investigated for Gram-positive bacteria. Previously, Bacillus velezensis LoaP was shown to enhance transcription of the difficidin antibiotic synthesis operon, but its antitermination determinants were unknown. In this study, we purified RNA polymerase and transcription factors, and reconstituted difficidin operon (dfn) antitermination activity in vitro. The dfn 5’ leader region contains a small RNA hairpin essential for antitermination in vivo and in vitro, and an intrinsic terminator that is highly dependent on NusA. The antitermination activity of LoaP emanates from its ability to specifically antagonize NusA; no other factors are required. These findings demonstrate the basic requirements of the LoaP class of NusG paralogs and indicate that the LoaP antitermination mechanism differs significantly from that of the RfaH paradigm.
HostingRepository	PRIDE
AnnounceDate	2025-10-09
AnnouncementXML	Submission_2025-10-09_12:45:23.681.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Conor Jenkins
SpeciesList	scientific name: Bacillus velezensis FZB42; NCBI TaxID: NEWT:326423; scientific name: Bacteria; NCBI TaxID: NCBITaxon:2;
ModificationList	phosphorylated residue; monohydroxylated residue; deaminated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2025-04-25 04:52:17	ID requested
⏵ 1	2025-10-09 12:45:24	announced

10.1128/MBIO.01429-25;

submitter keyword: transcription, LoaP,Bacillus, LCMS, Antitermination, Antibiotics

Wade Winkler
contact affiliation	The University of Maryland Department of Cell Biology & Molecular Genetics College Park, Maryland, USA
contact email	wwinkler@umd.edu
lab head
Conor Jenkins
contact affiliation	US Army DEVCOM CBC
contact email	conor.c.jenkins.civ@army.mil
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD063323
     1. Label: PRIDE project
     2. Name: Bacillus velezensis LoaP Promotes Antitermination of Antibiotic Synthesis Operons by Antagonizing NusA