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

Abstract

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 (MdbA Cd). Crystallization studies uncovered that the 1.2-Å resolution structure ofC. matruchotiiMdbA (MdbA Cm) 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 MdbA Cmcatalyzes disulfide bond formation within the actinobacterial pilin FimA. A new gene deletion method supported thatmdbAis essential inC. matruchotii. Remarkably, heterologous expression of MdbA Cmin 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 MdbA Cmis 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 biofilmsmore » 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.« less

Authors:
; ; ; ; ; ;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
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
OSTI Identifier:
1461493
DOE Contract Number:  
AC02-06CH11357
Resource Type:
Journal Article
Journal Name:
Journal of Bacteriology
Additional Journal Information:
Journal Volume: 200; Journal Issue: 9; Journal ID: ISSN 0021-9193
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English

Citation Formats

Luong, Truc Thanh, Tirgar, Reyhaneh, Reardon-Robinson, Melissa E., Joachimiak, Andrzej, Osipiuk, Jerzy, Ton-That, Hung, and Stock, Ann M. Structural Basis of a Thiol-Disulfide Oxidoreductase in the Hedgehog-Forming Actinobacterium Corynebacterium matruchotii. United States: N. p., 2018. Web. doi:10.1128/JB.00783-17.
Luong, Truc Thanh, Tirgar, Reyhaneh, Reardon-Robinson, Melissa E., Joachimiak, Andrzej, Osipiuk, Jerzy, Ton-That, Hung, & Stock, Ann M. Structural Basis of a Thiol-Disulfide Oxidoreductase in the Hedgehog-Forming Actinobacterium Corynebacterium matruchotii. United States. doi:10.1128/JB.00783-17.
Luong, Truc Thanh, Tirgar, Reyhaneh, Reardon-Robinson, Melissa E., Joachimiak, Andrzej, Osipiuk, Jerzy, Ton-That, Hung, and Stock, Ann M. Mon . "Structural Basis of a Thiol-Disulfide Oxidoreductase in the Hedgehog-Forming Actinobacterium Corynebacterium matruchotii". United States. doi:10.1128/JB.00783-17.
@article{osti_1461493,
title = {Structural Basis of a Thiol-Disulfide Oxidoreductase in the Hedgehog-Forming Actinobacterium Corynebacterium matruchotii},
author = {Luong, Truc Thanh and Tirgar, Reyhaneh and Reardon-Robinson, Melissa E. and Joachimiak, Andrzej and Osipiuk, Jerzy and Ton-That, Hung and Stock, Ann M.},
abstractNote = {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.},
doi = {10.1128/JB.00783-17},
journal = {Journal of Bacteriology},
issn = {0021-9193},
number = 9,
volume = 200,
place = {United States},
year = {2018},
month = {2}
}