Crystal structure of dehydratase component HadAB complex of mycobacterial FAS-II pathway

Biswas, Rupam; Dutta, Avijit; Dutta, Debajyoti; Hazra, Ditipriya; Banerjee, Deb Ranjan; Basak, Amit; Das, Amit Kumar

doi:10.1016/j.bbrc.2015.01.119

Cited by 17 publications

(27 citation statements)

References 29 publications

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“…This is further substantiated by comparison to the crystal structure of HadAB in the vicinity of the putative CoA moiety. By docking studies, it was proposed that HadAB can bind to both CoA and ACP, consistent with the activity HadAB exhibits toward both CoA‐ and ACP‐bound lipids (Biswas et al ., ). HadA Arg113 is involved in a salt‐bridge interaction with the CoA phosphate group (http://onlinelibrary.wiley.com/doi/10.1111/mmi.12992/suppinfo).…”

Section: Resultsmentioning

confidence: 97%

“…However, while HadA and HadC form an active enzyme by associating with the essential HadB component, it appears that MSMEG_6754 contains both components in a single protein. The structure of the M. smegmatis HadAB has recently been solved (Biswas et al, 2015). To determine the similarity of the structures, recombinant MSMEG_6754 was purified from Escherichia coli, crystallized, and its structure solved at 1.5 Å resolution by single anomalous dispersion, with refinement statistics as reported in Table S2.…”

Section: Expression Of Msmeg_6754 In M Tuberculosis Generates High Rmentioning

confidence: 99%

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A new dehydratase conferring innate resistance to thiacetazone and intra‐amoebal survival of Mycobacterium smegmatis

Carrère-Kremer

Blaise

Singh

et al. 2015

Molecular Microbiology

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SummaryNontuberculous mycobacteria are innately resistant to most antibiotics, although the mechanisms responsible for their drug resistance remain poorly understood. They are particularly refractory to thiacetazone (TAC), a second-line antitubercular drug. Herein, we identified MSMEG_6754 as essential for the innate resistance of Mycobacterium smegmatis to TAC. Transposon-mediated and targeted disruption of MSMEG_6754 resulted in hypersusceptibility to TAC. Conversely, introduction of MSMEG_6754 into Mycobacterium tuberculosis increased resistance 100-fold. Resolution of the crystal structure of MSMEG_6754 revealed a homodimer in which each monomer comprises two hot-dog domains characteristic of dehydratase-like proteins and very similar to the HadAB complex involved in mycolic acid biosynthesis. Gene inactivation of the essential hadB dehydratase could be achieved in M. smegmatis and M. tuberculosis only when the strains carried an integrated copy of MSMEG_6754, supporting the idea that MSMEG_6754 and HadB share redundant dehydratase activity. Using M. smegmatis-Acanthamoeba co-cultures, we found that intra-amoebal growth of the MSMEG_6754 deleted strain was significantly reduced compared with the parental strain. This in vivo growth defect was fully restored upon complementation with catalytically active MSMEG_6754 or HadABC, indicating that MSMEG_6754 plays a critical role in the survival of M. smegmatis within the environmental host.

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Section: Resultsmentioning

confidence: 97%

Section: Expression Of Msmeg_6754 In M Tuberculosis Generates High Rmentioning

confidence: 99%

A new dehydratase conferring innate resistance to thiacetazone and intra‐amoebal survival of Mycobacterium smegmatis

Carrère-Kremer

Blaise

Singh

et al. 2015

Molecular Microbiology

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“…The (3 R )-hydroxyacyl-ACP dehydratases HadAB and HadBC are two main components of the mycobacterial FAS-II system, HadA and HadC being the substrate-binding subunits and HadB the catalytic subunit 21 , 22 . Accordingly, HadB is essential for the MA biosynthesis in M .…”

Section: Resultsmentioning

confidence: 99%

HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness

Lefebvre

Boulon

Ducoux

et al. 2018

Sci Rep

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Mycolic acids (MAs) have a strategic location within the mycobacterial envelope, deeply influencing its architecture and permeability, and play a determinant role in the pathogenicity of mycobacteria. The fatty acid synthase type II (FAS-II) multienzyme system is involved in their biosynthesis. A combination of pull-downs and proteomics analyses led to the discovery of a mycobacterial protein, HadD, displaying highly specific interactions with the dehydratase HadAB of FAS-II. In vitro activity assays and homology modeling showed that HadD is, like HadAB, a hot dog folded (R)-specific hydratase/dehydratase. A hadD knockout mutant of Mycobacterium smegmatis produced only the medium-size alpha’-MAs. Data strongly suggest that HadD is involved in building the third meromycolic segment during the late FAS-II elongation cycles, leading to the synthesis of the full-size alpha- and epoxy-MAs. The change in the envelope composition induced by hadD inactivation strongly altered the bacterial fitness and capacities to aggregate, assemble into colonies or biofilms and spread by sliding motility, and conferred a hypersensitivity to the firstline antimycobacterial drug rifampicin. This showed that the cell surface properties and the envelope integrity were greatly affected. With the alarmingly increasing case number of nontuberculous mycobacterial diseases, HadD appears as an attractive target for drug development.

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“…The essential dehydratase step in the biosynthesis of MAs is performed by the HadA-HadB and HadB-HadC complexes [ 14 , 15 ]. The HadB subunit bears the catalytic domain whereas HadA and HadC determine the substrate specificity [ 14 , 47 ]. In this work we showed, that in M .…”

Section: Discussionmentioning

confidence: 99%

The Non-Essential Mycolic Acid Biosynthesis Genes hadA and hadC Contribute to the Physiology and Fitness of Mycobacterium smegmatis

Jamet¹,

Slama²,

Domingues³

et al. 2015

PLoS ONE

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Gram positive mycobacteria with a high GC content, such as the etiological agent of tuberculosis Mycobacterium tuberculosis, possess an outer membrane mainly composed of mycolic acids (MAs), the so-called mycomembrane, which is essential for the cell. About thirty genes are involved in the biosynthesis of MAs, which include the hadA, hadB and hadC genes that encode the dehydratases Fatty Acid Synthase type II (FAS-II) known to function as the heterodimers HadA-HadB and HadB-HadC. The present study shows that M. smegmatis cells remain viable in the absence of either HadA and HadC or both. Inactivation of HadC has a dramatic effect on the physiology and fitness of the mutant strains whereas that of HadA exacerbates the phenotype of a hadC deletion. The hadC mutants exhibit a novel MA profile, display a distinct colony morphology, are less aggregated, are impaired for sliding motility and biofilm development and are more resistant to detergent. Conversely, the hadC mutants are significantly more susceptible to low- and high-temperature and to selective toxic compounds, including several current anti-tubercular drugs.

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Crystal structure of dehydratase component HadAB complex of mycobacterial FAS-II pathway

Cited by 17 publications

References 29 publications

A new dehydratase conferring innate resistance to thiacetazone and intra‐amoebal survival of Mycobacterium smegmatis

A new dehydratase conferring innate resistance to thiacetazone and intra‐amoebal survival of Mycobacterium smegmatis

HadD, a novel fatty acid synthase type II protein, is essential for alpha- and epoxy-mycolic acid biosynthesis and mycobacterial fitness

The Non-Essential Mycolic Acid Biosynthesis Genes hadA and hadC Contribute to the Physiology and Fitness of Mycobacterium smegmatis

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