Biochemical, structural, and functional studies reveal that MAB_4324c from <i>Mycobacterium abscessus</i> is an active tandem repeat <i>N</i>‐acetyltransferase

Alsarraf, Husam M. A. B.; Ung, Kien Lam; Johansen, Matt D.; Dimon, Juliette; Oliéric, V.; Kremer, Laurent; Blaise, Mickaël

doi:10.1002/1873-3468.14360

Cited by 3 publications

(3 citation statements)

References 68 publications

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“…Additionally, we identified two genes, MAB_2142 and MAB_2146, activated only in Mabs-R by Lsr2 and encoding [4F-4S] Ferredoxin proteins that function as intracellular stress sensors and contribute to genomic stability during macrophage and amoeba infections ( 30 ). Deletion of lsr2 in Mabs-S and Mabs-R also results in a significant decrease in the expression of the MAB_4324 c gene, which encodes an N-acetyltransferase recently found to enhance internalization and intracellular growth of mycobacteria in human macrophages ( 31 ) ( Fig. 2A ).…”

Section: Resultsmentioning

confidence: 86%

Lsr2, a pleiotropic regulator at the core of the infectious strategy of Mycobacterium abscessus

Gerges,

Rodríguez-Ordoñez,

Durand

et al. 2024

Microbiol Spectr

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Mycobacterium abscessus is a non-tuberculous mycobacterium, causing lung infections in cystic fibrosis patients. During pulmonary infection, M. abscessus switches from smooth (Mabs-S) to rough (Mabs-R) morphotypes, the latter being hyper-virulent. Previously, we isolated the lsr2 gene as differentially expressed during S-to-R transition. lsr2 encodes a pleiotropic transcription factor that falls under the superfamily of nucleoid-associated proteins. Here, we used two functional genomic methods, RNA-seq and chromatin immunoprecipitation-sequencing (ChIP-seq), to elucidate the molecular role of Lsr2 in the pathobiology of M. abscessus . Transcriptomic analysis shows that Lsr2 differentially regulates gene expression across both morphotypes, most of which are involved in several key cellular processes of M. abscessus , including host adaptation and antibiotic resistance. These results were confirmed through quantitative real-time PCR, as well as by minimum inhibitory concentration tests and infection tests on macrophages in the presence of antibiotics. ChIP-seq analysis revealed that Lsr2 extensively binds the M. abscessus genome at AT-rich sequences and appears to form long domains that participate in the repression of its target genes. Unexpectedly, the genomic distribution of Lsr2 revealed no distinctions between Mabs-S and Mabs-R, implying more intricate mechanisms at play for achieving target selectivity. IMPORTANCE Lsr2 is a crucial transcription factor and chromosome organizer involved in intracellular growth and virulence in the smooth and rough morphotypes of Mycobacterium abscessus . Using RNA-seq and chromatin immunoprecipitation-sequencing (ChIP-seq), we investigated the molecular role of Lsr2 in gene expression regulation along with its distribution on M. abscessus genome. Our study demonstrates the pleiotropic regulatory role of Lsr2, regulating the expression of many genes coordinating essential cellular and molecular processes in both morphotypes. In addition, we have elucidated the role of Lsr2 in antibiotic resistance both in vitro and in vivo , where lsr2 mutant strains display heightened sensitivity to antibiotics. Through ChIP-seq, we reported the widespread distribution of Lsr2 on M. abscessus genome, revealing a direct repressive effect due to its extensive binding on promoters or coding sequences of its targets. This study unveils the significant regulatory role of Lsr2, intricately intertwined with its function in shaping the organization of the M. abscessus genome.

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Section: Resultsmentioning

confidence: 86%

Lsr2, a pleiotropic regulator at the core of the infectious strategy of Mycobacterium abscessus

Gerges,

Rodríguez-Ordoñez,

Durand

et al. 2024

Microbiol Spectr

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show abstract

“…Additionally, we identified two genes, MAB_2142 and MAB_2146 activated only in Mabs-R by Lsr2 and encoding [4F-4S] Ferredoxin proteins that function as intracellular stress sensors and contribute to genomic stability during macrophage and amoeba infections (30). Deletion of lsr2 in Mabs-S and Mabs-R also results in a significant decrease in the expression of the MAB_4324c gene, which encodes an N-acetyltransferase recently found to enhance internalization and intracellular growth of mycobacteria in human macrophages (31) ( Figure 2A ).…”

Section: Resultsmentioning

confidence: 90%

Lsr2, a pleiotropic regulator at the core of the infectious strategy ofMycobacterium abscessus

Gerges,

del Pilar Rodríguez-Ordoñez,

Durand

et al. 2023

Preprint

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Mycobacterium abscessusis a non-tuberculous mycobacterium, causing lung infections in cystic fibrosis patients. During pulmonary infection,M. abscessusswitches from smooth (Mabs-S) to rough (Mabs-R) morphotypes, the latter being hyper-virulent. Previously, we isolated thelsr2gene as differentially expressed during S-to-R transition.lsr2encodes a pleiotropic transcription factor that falls under the superfamily of nucleoid-associated proteins (NAPs). Here, we used two functional genomics methods, RNA-seq and ChIP-seq, to elucidate the molecular role of Lsr2 in the pathobiology ofM. abscessus. Transcriptomic analysis shows that Lsr2 differentially regulates gene expression across both morphotypes, most of which are involved in several key cellular processes ofM. abscessus, including host adaptation and antibiotic resistance. These results were confirmed through RT-qPCR, as well as by minimum inhibitory concentration (MIC) tests and infection tests on macrophages in the presence of antibiotics. ChIP-seq analysis revealed that Lsr2 extensively binds theM. abscessusgenome at AT-rich sequences and appears to form long domains that participate in the repression of its target genes. Unexpectedly, the genomic distribution of Lsr2 revealed no distinctions between Mabs-S and Mabs-R, implying more intricate mechanisms at play for achieving target selectivity.IMPORTANCELsr2 is a crucial transcription factor and chromosome organizer involved in intracellular growth and virulence in the smooth and rough morphotypes ofMycobacterium abscessus(Mabs). Using RNA-seq and ChIP-seq, we investigated the molecular role of Lsr2 in gene expression regulation along with its distribution on Mabs genome. Our study demonstrates the pleiotropic regulatory role of Lsr2, regulating the expression of many genes coordinating essential cellular and molecular processes in both morphotypes. In addition, we have elucidated the role of Lsr2 in antibiotic resistance bothin vitroandin vivo, wherelsr2mutant strains display heightened sensitivity to antibiotics. Through ChIP-seq, we reported the widespread distribution of Lsr2 on Mabs genome, revealing a direct repressive effect due to its extensive binding on promoters or coding sequences of its targets. This study unveils the significant regulatory role of Lsr2, intricately intertwined with its function in shaping the organization of the Mabs genome.

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“…We tested 4 acetyltransferases with unknown functions and a phosphotransferase of which MAB_4324c showed only a modest ability to complement the AMK sensitivity of Δ MabwhiB7 (Figs 3B and S2A–S2D ). Interestingly, the AMK sensitivity of a ΔMAB_4324c deletion strain was previously shown to be indistinguishable from that of WT bacteria; purified Mab4324c nonetheless showed mild acetylation of AMK [ 36 ]. These results are consistent with the modest AMK resistance conferred by MAB_4324c in the AMK hypersensitive background of Δ MabwhiB7 and is suggestive of a relatively minor role of MAB_4324c in AMK resistance of M .…”

Section: Resultsmentioning

confidence: 99%

Hierarchy and interconnected networks in the WhiB7 mediated transcriptional response to antibiotic stress in Mycobacterium abscessus

Hurst-Hess,

McManaman,

Yang

et al. 2023

PLoS Genet

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Mycobacterium abscessus is intrinsically resistant to antibiotics effective against other pathogenic mycobacteria largely due to the drug-induced expression of genes that confer resistance. WhiB7 is a major hub controlling the induction of resistance to ribosome-targeting antibiotics. It activates the expression of >100 genes, 7 of which are known determinants of drug resistance; the function of most genes within the regulon is however unknown, but some conceivably encode additional mechanisms of resistance. Furthermore, the hierarchy of gene expression within the regulon, if any, is poorly understood. In the present work we have identified 56 WhiB7 binding sites using chromatin immunoprecipitation sequencing (CHIP-Seq) which accounts for the WhiB7-dependent upregulation of 72 genes, and find that M. abscessus WhiB7 functions exclusively as a transcriptional activator at promoters recognized by σA/σB. We have investigated the role of 18 WhiB7 regulated genes in drug resistance. Our results suggest that while some genes within the regulon (eg. erm41, hflX, eis2 and the ABCFs) play a major role in resistance, others make smaller contributions (eg. MAB_4324c and MAB_1409c) and the observed hypersensitivity ΔMabwhiB7 is a cumulative effect of these individual contributions. Moreover, our CHIP-Seq data implicate additional roles of WhiB7 induced genes beyond antibiotic resistance. Finally, we identify a σH-dependent network in aminoglycoside and tigecycline resistance which is induced upon drug exposure and is further activated by WhiB7 demonstrating the existence of a crosstalk between components of the WhiB7-dependent and -independent circuits.

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Biochemical, structural, and functional studies reveal that MAB_4324c from Mycobacterium abscessus is an active tandem repeat N‐acetyltransferase

Cited by 3 publications

References 68 publications

Lsr2, a pleiotropic regulator at the core of the infectious strategy of Mycobacterium abscessus

Lsr2, a pleiotropic regulator at the core of the infectious strategy of Mycobacterium abscessus

Lsr2, a pleiotropic regulator at the core of the infectious strategy ofMycobacterium abscessus

Hierarchy and interconnected networks in the WhiB7 mediated transcriptional response to antibiotic stress in Mycobacterium abscessus

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