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Structural Basis of a Thiol-Disulfide Oxidoreductase in the Hedgehog-Forming Actinobacterium Corynebacterium matruchotii

Journal Article · · Journal of Bacteriology
DOI:https://doi.org/10.1128/JB.00783-17· OSTI ID:1461493
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ABSTRACT

The actinobacteriumCorynebacterium matruchotiihas been implicated in nucleation of oral microbial consortia leading to biofilm formation. Due to the lack of genetic tools, little is known about basic cellular processes, including protein secretion and folding, in this organism. We report here a survey of theC. matruchotiigenome, which encodes a large number of exported proteins containing paired cysteine residues, and identified an oxidoreductase that is highly homologous to theCorynebacterium diphtheriaethiol-disulfide oxidoreductase MdbA (MdbACd). Crystallization studies uncovered that the 1.2-Å resolution structure ofC. matruchotiiMdbA (MdbACm) possesses two conserved features found in actinobacterial MdbA enzymes, a thioredoxin-like fold and an extended α-helical domain. By reconstituting the disulfide bond-forming machinein vitro, we demonstrated that MdbACmcatalyzes disulfide bond formation within the actinobacterial pilin FimA. A new gene deletion method supported thatmdbAis essential inC. matruchotii. Remarkably, heterologous expression of MdbACmin theC. diphtheriaeΔmdbAmutant rescued its known defects in cell growth and morphology, toxin production, and pilus assembly, and this thiol-disulfide oxidoreductase activity required the catalytic motif CXXC. Altogether, the results suggest that MdbACmis a major thiol-disulfide oxidoreductase, which likely mediates posttranslocational protein folding inC. matruchotiiby a mechanism that is conserved inActinobacteria.

IMPORTANCEThe actinobacteriumCorynebacterium matruchotiihas been implicated in the development of oral biofilms or dental plaque; however, little is known about the basic cellular processes in this organism. We report here a high-resolution structure of aC. matruchotiioxidoreductase that is highly homologous to theCorynebacterium diphtheriaethiol-disulfide oxidoreductase MdbA. By biochemical analysis, we demonstrated thatC. matruchotiiMdbA catalyzes disulfide bond formationin vitro. Furthermore, a new gene deletion method revealed that deletion ofmdbAis lethal inC. matruchotii. Remarkably,C. matruchotiiMdbA can replaceC. diphtheriaeMdbA to maintain normal cell growth and morphology, toxin production, and pilus assembly. Overall, our studies support the hypothesis thatC. matruchotiiutilizes MdbA as a major oxidoreductase to catalyze oxidative protein folding.

Research Organization:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science - Office of Biological and Environmental Research; National Institutes of Health (NIH) - National Institute of Allergy and Infectious Diseases (NIAID); National Institutes of Health (NIH) - National Institute of Dental and Craniofacial Research
DOE Contract Number:
AC02-06CH11357
OSTI ID:
1461493
Journal Information:
Journal of Bacteriology, Vol. 200, Issue 9; ISSN 0021-9193
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English

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