Induced DNA bending by unique dimerization of HigA antitoxin

Park, Jin‐Young; Kim, Hyo Jung; Pathak, Chinar; Yoon, Hye‐Jin; Kim, Do-Hee; Park, Sung Jean; Lee, Bong-Jin

doi:10.1107/s2052252520006466

Cited by 7 publications

(7 citation statements)

References 51 publications

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“…Recently, CBU1490 was found to be structurally homologous to the Mycobacterium tuberculosis HigA3 antitoxin (Park et al, 2020). Based on the finding of another potential antitoxin in C. burnetii, we mined the genome for other putative toxin and antitoxins.…”

Section: Previous Attempts To Use the Dcas9 From Staphylococcus Aureu...mentioning

confidence: 99%

The endogenous Coxiella burnetii plasmid encodes a functional toxin–antitoxin system

Wachter

Cockrell

Miller³

et al. 2022

Molecular Microbiology

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Coxiella burnetii is the causative agent of Q fever. All C. burnetii isolates encode either an autonomously replicating plasmid (QpH1, QpDG, QpRS, or QpDV) or QpRS-like chromosomally integrated plasmid sequences. The role of the ORFs present in these sequences is unknown. Here, the role of the ORFs encoded on QpH1 was investigated.Using a new C. burnetii shuttle vector (pB-TyrB-QpH1ori), we cured the C. burnetii Nine Mile Phase II strain of QpH1. The ΔQpH1 strain grew normally in axenic media but had a significant growth defect in Vero cells, indicating QpH1 was important for C. burnetii virulence. We developed an inducible CRISPR interference system to examine the role of individual QpH1 plasmid genes. CRISPRi of cbuA0027 resulted in significant growth defects in axenic media and THP-1 cells. The cbuA0028/cbuA0027 operon encodes CBUA0028 (ToxP) and CBUA0027 (AntitoxP), which are homologous to the HigB2 toxin and HigA2 antitoxin, respectively, from Vibrio cholerae. Consistent with toxin-antitoxin systems, overexpression of toxP resulted in a severe intracellular growth defect that was rescued by co-expression of antitoxP. ToxP inhibited protein translation. AntitoxP bound the toxP promoter (PtoxP) and ToxP, with the resulting complex binding also PtoxP. In summary, our data indicate that C. burnetii maintains an autonomously replicating plasmid because of a plasmid-based toxin-antitoxin system.

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Section: Previous Attempts To Use the Dcas9 From Staphylococcus Aureu...mentioning

confidence: 99%

The endogenous Coxiella burnetii plasmid encodes a functional toxin–antitoxin system

Wachter

Cockrell

Miller³

et al. 2022

Molecular Microbiology

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“…The canonical hierarchy for TA systems has the antitoxin gene located upstream of the toxin, likely as a regulatory control, but in the HigBA system this gene order is swapped. It is not known why the HigB toxin gene is the first gene of the operon, although the antitoxin does not possess its own promoter (Armalytė et al, 2018;Park et al, 2020). However, unlike the previous study on Mt HigBA3, the promoter region for Mt HigBA2 was not clearly defined.…”

Section: The Unique Characteristics Of M Tuberculosis Higa2mentioning

confidence: 83%

“…of 2.6 Å ), Mt HigA2 shows a major difference in the arrangement of the two HTH motifs from each monomer. Mt HigA3 showed an arched formation of HTH motifs, while Mt HigA2 has a linear arrangement (Park et al, 2020). This linear configuration is unique compared with similar transcription factors [Fig.…”

Section: The Unique Characteristics Of M Tuberculosis Higa2mentioning

confidence: 99%

Chasing the structural diversity of the transcription regulator Mycobacterium tuberculosis HigA2

Richardson

Kang

Lee

et al. 2021

IUCrJ

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Transcription factors are the primary regulators of gene expression and recognize specific DNA sequences under diverse physiological conditions. Although they are vital for many important cellular processes, it remains unclear when and how transcription factors and DNA interact. The antitoxin from a toxin–antitoxin system is an example of negative transcriptional autoregulation: during expression of the cognate toxin it is suppressed through binding to a specific DNA sequence. In the present study, the antitoxin HigA2 from Mycobacterium tuberculosis M37Rv was structurally examined. The crystal structure of M. tuberculosis HigA2 comprises three sections: an N-terminal autocleavage region, an α-helix bundle which contains an HTH motif, and a C-terminal β-lid. The N-terminal region is responsible for toxin binding, but was shown to cleave spontaneously in its absence. The HTH motif performs a key role in DNA binding, with the C-terminal β-lid influencing the interaction by mediating the distance between the motifs. However, M. tuberculosis HigA2 exhibits a unique coordination of the HTH motif and no DNA-binding activity is detected. Three crystal structures of M. tuberculosis HigA2 show a flexible alignment of the HTH motif, which implies that the motif undergoes structural rearrangement to interact with DNA. This study reveals the molecular mechanisms of how transcription factors interact with partner proteins or DNA.

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“…CBUA0028 displays 26% identity with HigB2 and conservation of the five amino acids (K49, R51, R64, Y82 and K84) previously shown to comprise the HigB2 active site (Hadzi et al, 2017). CBUA0027 has 30% identity with HigA2 and displays a conserved helix-turn helix motif that is essential for DNA binding (Park et al, 2020). These data suggested CBUA0028 and CBUA0027 function as a toxin and antitoxin, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…CBU1490 is identified as an addiction molecule antidote protein and its crystal structure has been solved (Franklin et al, 2015). Recently CBU1490 was found to be structurally homologous to the Mycobacterium tuberculosis HigA3 antitoxin (Park et al, 2020). Based on the finding of another potential antitoxin in C. burnetii we mined the genome for other putative toxin and antitoxins.…”

Section: Discussionmentioning

confidence: 99%

A toxin-antitoxin system ensures plasmid stability in Coxiella burnetii

Wachter¹,

Cockrell

Miller³

et al. 2022

Preprint

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Coxiella burnetii is the causative agent of Q fever. All C. burnetii isolates encode either an autonomous replicating plasmid (QpH1, QpDG, QpRS, or QpDV) or QpRS-like chromosomally integrated plasmid sequences. The role of the ORFs present on these sequences is unknown. Here, the role of the ORFs encoded on QpH1 was investigated. Using a new C. burnetii shuttle vector (pB-TyrB-QpH1ori) we cured Nine Mile Phase II of QpH1. The ΔQpH1 strain grew normally in axenic media but had a significant growth defect in Vero cells, indicating QpH1 was important for C. burnetii virulence. We developed an inducible CRISPR interference system to examine the role of individual QpH1 plasmid genes. CRISPRi of cbuA0027 resulted in significant growth defects in axenic media and THP-1 cells. The cbuA0028/cbuA0027 operon encodes CBUA0028 and CBUA0027, which are homologous to the HigB2 toxin and HigA2 anti-toxin, respectively, from Vibrio cholerae. Consistent with toxin-antitoxin systems, overexpression of cbuA0028 resulted in a severe intracellular growth defect that was rescued by co-expression of cbuA0027. CBUA0028 inhibited protein translation. CBUA0027 bound the cbuA0028 promoter (PcbuA0028) and CBUA0028, with the resulting complex binding also PcbuA0028. In summary, our data indicates C. burnetii maintains an autonomously replicating plasmid because of a plasmid-based toxin-antitoxin system.

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Induced DNA bending by unique dimerization of HigA antitoxin

Cited by 7 publications

References 51 publications

The endogenous Coxiella burnetii plasmid encodes a functional toxin–antitoxin system

The endogenous Coxiella burnetii plasmid encodes a functional toxin–antitoxin system

Chasing the structural diversity of the transcription regulator Mycobacterium tuberculosis HigA2

A toxin-antitoxin system ensures plasmid stability in Coxiella burnetii

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