PXD046632-2

PXD046632 is an original dataset announced via ProteomeXchange.

Title	Molecular insights into RNA polymerase recycling by mycobacterial HelD
Description	Mycobacterial HelD is a transcription factor that binds RNA polymerase (RNAP) and thereby rescues it from stalled transcription complexes. HelD also protects RNAP against the antibiotic rifampicin. HelD, however, must be released from the rescued RNAP to participate in the next round of the transcription cycle. The exact mechanism of this release is unknown. Here we show that HelD from Mycobacterium smegmatis can form a complex with RNAP, together with σA primary sigma factor, and RbpA, but not CarD, transcription factors. Using cryo-EM, we solved a series of RNAP-σA-RbpA-HelD structures with or without promoter DNA. These snapshots capture the involvement of HelD in transcription initiation, describes mechanistic aspects of HelD release, and its protective effect against rifampicin. Biochemical evidence supports these findings, defines the role of ATP binding/hydrolysis to/by HelD in the process, and functionally confirms the rifampicin-protective effect of HelD. Taken together, HelD assists an alternative pathway of transcription initiation.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_07:00:53.655.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD046632
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Alena Krenkova
SpeciesList	scientific name: Mycobacterium smegmatis; NCBI TaxID: 1772;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-11-03 03:05:12	ID requested
1	2024-10-08 03:41:55	announced
⏵ 2	2024-10-22 07:00:54	announced	2024-10-22: Updated project metadata.

10.1038/S41467-024-52891-5;

10.6019/PXD046632;

submitter keyword: HelD

RNA polymerase

Mycobacteria

Transcription

Rifampicin

Martin Hubalek
contact affiliation	IOCB CAS CR
contact email	hubalek@uochb.cas.cz
lab head
Alena Krenkova
contact affiliation	IOCB CAS CR
contact email	alena.krenkova@uochb.cas.cz
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD046632
     1. Label: PRIDE project
     2. Name: Molecular insights into RNA polymerase recycling by mycobacterial HelD