Control of replication and gene expression by ADP-ribosylation of DNA in Mycobacterium tuberculosis

Journal article

Mycobacterium tuberculosis maintains long-term infections characterised by the need to regulate growth and adapt to contrasting in vivo environments. Here we show that M. tuberculosis complex bacteria utilise reversible ADP-ribosylation of single-stranded DNA as a mechanism to coordinate stationary phase growth with transcriptional adaptation. The DNA modification is controlled by DarT, an ADP-ribosyltransferase, which adds ADP-ribose to thymidine, and DarG, which enzymatically removes this base modification. Using darG-knockdown M. bovis BCG, we map the first DNA ADP-ribosylome from any organism. We show that inhibition of replication by DarT is reversible and accompanied by extensive ADP-ribosylation at the origin of replication (OriC). In addition, we observe ADP-ribosylation across the genome and demonstrate that ADP-ribose-thymidine alters the transcriptional activity of M. tuberculosis RNA polymerase. Furthermore, we demonstrate that during stationary phase, DarT-dependent ADP-ribosylation of M. tuberculosis DNA is required to optimally induce expression of the Zur regulon, including the ESX-3 secretion system and multiple alternative ribosome proteins. Thus, ADP-ribosylation of DNA can provide a mechanistic link through every aspect of DNA biology from replication to transcription to translation.

Authors: Butler, RE; Schuller, M; Jaiswal, R; Mukhopadhyay, J; Barber, J

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: EMBO Press
• Journal: The EMBO Journal
• Volume: 44
• Issue: 12
• Pages: 3468-3491
• Publication date: 2025-05-08
• Acceptance date: 2025-04-22
• DOI: 10.1038/s44318-025-00451-y
• EISSN: 1460-2075
• ISSN: 0261-4189
• Language: English
• Keywords: PARP; ADP-ribosylation; ADPr-Seq; Transcription Regulation; DNA Modification