Mycobacterium smegmatis PafBC is involved in regulation of DNA damage response

Scientific Reports
Begonia Fudrini OlivenciaFrank Imkamp

Abstract

Two genes, pafB and pafC, are organized in an operon with the Pup-ligase gene pafA, which is part of the Pup-proteasome system (PPS) present in mycobacteria and other actinobacteria. The PPS is crucial for Mycobacterium tuberculosis resistance towards reactive nitrogen intermediates (RNI). However, pafB and pafC apparently play only a minor role in RNI resistance. To characterize their function, we generated a pafBC deletion in Mycobacterium smegmatis (Msm). Proteome analysis of the mutant strain revealed decreased cellular levels of various proteins involved in DNA damage repair, including recombinase A (RecA). In agreement with this finding, Msm ΔpafBC displayed increased sensitivity to DNA damaging agents. In mycobacteria two pathways regulate DNA repair genes: the LexA/RecA-dependent SOS response and a predominant pathway that controls gene expression via a LexA/RecA-independent promoter, termed P1. PafB and PafC feature winged helix-turn-helix DNA binding motifs and we demonstrate that together they form a stable heterodimer in vitro, implying a function as a heterodimeric transcriptional regulator. Indeed, P1-driven transcription of recA was decreased in Msm ΔpafBC under standard conditions and induction of recA expressio...Continue Reading

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Citations

Jan 12, 2020·Current Topics in Medicinal Chemistry·Pooja MittalIndrakant K Singh
Apr 27, 2019·Frontiers in Molecular Biosciences·Andreas U Müller, Eilika Weber-Ban
Jan 21, 2021·Antimicrobial Agents and Chemotherapy·Marissa B GuzzoLiem Nguyen
Jun 29, 2021·Frontiers in Molecular Biosciences·Tatjana von RosenEilika Weber-Ban

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Methods Mentioned

BETA
ubiquitination
gel filtration
PCR
electrophoretic mobility shift assay
acetylation

Software Mentioned

BLAST
Primer3

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