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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}
}

Works referenced in this record:

Identification of a protein required for disulfide bond formation in vivo
journal, November 1991


MOLPROBITY: structure validation and all-atom contact analysis for nucleic acids and their complexes
journal, July 2004

  • Davis, I. W.; Murray, L. W.; Richardson, J. S.
  • Nucleic Acids Research, Vol. 32, Issue Web Server
  • DOI: 10.1093/nar/gkh398

Requirement of Signal Peptidase ComC and Thiol-Disulfide Oxidoreductase DsbA for Optimal Cell Surface Display of Pseudopilin ComGC in Staphylococcus aureus
journal, July 2012

  • van der Kooi-Pol, Magdalena M.; Reilman, Ewoud; Sibbald, Mark J. J. B.
  • Applied and Environmental Microbiology, Vol. 78, Issue 19
  • DOI: 10.1128/AEM.01565-12

Sortases and pilin elements involved in pilus assembly of Corynebacterium diphtheriae: Pilus assembly in corynebacteria
journal, May 2004


Refinement of Macromolecular Structures by the Maximum-Likelihood Method
journal, May 1997

  • Murshudov, G. N.; Vagin, A. A.; Dodson, E. J.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 53, Issue 3
  • DOI: 10.1107/S0907444996012255

Biogeography of a human oral microbiome at the micron scale
journal, January 2016

  • Mark Welch, Jessica L.; Rossetti, Blair J.; Rieken, Christopher W.
  • Proceedings of the National Academy of Sciences, Vol. 113, Issue 6
  • DOI: 10.1073/pnas.1522149113

Protein export through the bacterial Sec pathway
journal, November 2016

  • Tsirigotaki, Alexandra; De Geyter, Jozefien; Šoštaric´, Nikolina
  • Nature Reviews Microbiology, Vol. 15, Issue 1
  • DOI: 10.1038/nrmicro.2016.161

The bdbDC Operon of Bacillus subtilis Encodes Thiol-disulfide Oxidoreductases Required for Competence Development
journal, December 2001

  • Meima, Rob; Eschevins, Caroline; Fillinger, Sabine
  • Journal of Biological Chemistry, Vol. 277, Issue 9
  • DOI: 10.1074/jbc.M111380200

Rv2969c, essential for optimal growth in Mycobacterium tuberculosis , is a DsbA-like enzyme that interacts with VKOR-derived peptides and has atypical features of DsbA-like disulfide oxidases
journal, September 2013

  • Premkumar, Lakshmanane; Heras, Begoña; Duprez, Wilko
  • Acta Crystallographica Section D Biological Crystallography, Vol. 69, Issue 10
  • DOI: 10.1107/S0907444913017800

Dali server: conservation mapping in 3D
journal, May 2010

  • Holm, Liisa; Rosenstr�m, P�ivi
  • Nucleic Acids Research, Vol. 38, Issue suppl_2
  • DOI: 10.1093/nar/gkq366

Sec-secretion and sortase-mediated anchoring of proteins in Gram-positive bacteria
journal, August 2014

  • Schneewind, Olaf; Missiakas, Dominique
  • Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, Vol. 1843, Issue 8
  • DOI: 10.1016/j.bbamcr.2013.11.009

Functional Analysis of Paralogous Thiol-disulfide Oxidoreductases in Streptococcus gordonii
journal, April 2013

  • Davey, Lauren; Ng, Crystal K. W.; Halperin, Scott A.
  • Journal of Biological Chemistry, Vol. 288, Issue 23
  • DOI: 10.1074/jbc.M113.464578

Coot model-building tools for molecular graphics
journal, November 2004

  • Emsley, Paul; Cowtan, Kevin
  • Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12, p. 2126-2132
  • DOI: 10.1107/S0907444904019158

Lethality of sortase depletion in A ctinomyces oris caused by excessive membrane accumulation of a surface glycoprotein
journal, September 2014

  • Wu, Chenggang; Huang, I‐Hsiu; Chang, Chungyu
  • Molecular Microbiology, Vol. 94, Issue 6
  • DOI: 10.1111/mmi.12780

A Disulfide Bond-forming Machine Is Linked to the Sortase-mediated Pilus Assembly Pathway in the Gram-positive Bacterium Actinomyces oris
journal, July 2015

  • Reardon-Robinson, Melissa E.; Osipiuk, Jerzy; Chang, Chungyu
  • Journal of Biological Chemistry, Vol. 290, Issue 35
  • DOI: 10.1074/jbc.M115.672253

Structural and biochemical characterization of the essential DsbA-like disulfide bond forming protein from Mycobacterium tuberculosis
journal, January 2013

  • Chim, Nicholas; Harmston, Christine A.; Guzman, David J.
  • BMC Structural Biology, Vol. 13, Issue 1
  • DOI: 10.1186/1472-6807-13-23

Effect of a Mutagenic Agent on Calcifiability of Bacterionema matruchotii
journal, July 1972


Disulfide-Bond-Forming Pathways in Gram-Positive Bacteria
journal, December 2015

  • Reardon-Robinson, Melissa E.; Ton-That, Hung
  • Journal of Bacteriology, Vol. 198, Issue 5
  • DOI: 10.1128/JB.00769-15

Thiol-Disulfide Exchange in Gram-Positive Firmicutes
journal, November 2016


PHENIX : building new software for automated crystallographic structure determination
journal, October 2002

  • Adams, Paul D.; Grosse-Kunstleve, Ralf W.; Hung, Li-Wei
  • Acta Crystallographica Section D Biological Crystallography, Vol. 58, Issue 11
  • DOI: 10.1107/S0907444902016657

A special relationship between spherical and filamentous microorganisms in mature human dental plaque
journal, March 1972


A pathway for disulfide bond formation in vivo.
journal, February 1993

  • Bardwell, J. C.; Lee, J. O.; Jander, G.
  • Proceedings of the National Academy of Sciences, Vol. 90, Issue 3
  • DOI: 10.1073/pnas.90.3.1038

Structure and Function of DsbA, a Key Bacterial Oxidative Folding Catalyst
journal, May 2011

  • Shouldice, Stephen R.; Heras, Begoña; Walden, Patricia M.
  • Antioxidants & Redox Signaling, Vol. 14, Issue 9
  • DOI: 10.1089/ars.2010.3344

Disulfide Bond Formation and Cysteine Exclusion in Gram-positive Bacteria
journal, November 2009

  • Daniels, Robert; Mellroth, Peter; Bernsel, Andreas
  • Journal of Biological Chemistry, Vol. 285, Issue 5
  • DOI: 10.1074/jbc.M109.081398

Genes required for mycobacterial growth defined by high density mutagenesis: Genes required for mycobacterial growth
journal, March 2003


Transposon-5 mutagenesis transforms Corynebacterium matruchotii to synthesize novel hybrid fatty acids that functionally replace corynomycolic acid
journal, July 2003

  • Takayama, Kuni; Hayes, Barry; Vestling, Martha M.
  • Biochemical Journal, Vol. 373, Issue 2
  • DOI: 10.1042/bj20030248

Calcification of a Cariogenic Streptococcus and of Corynebacterium (Bacterionema) matruchotii
journal, June 1993


The Corynebacterium diphtheriae shaft pilin SpaA is built of tandem Ig-like modules with stabilizing isopeptide and disulfide bonds
journal, September 2009

  • Kang, H. J.; Paterson, N. G.; Gaspar, A. H.
  • Proceedings of the National Academy of Sciences, Vol. 106, Issue 40
  • DOI: 10.1073/pnas.0906826106

Reoxidation of the Thiol-Disulfide Oxidoreductase MdbA by a Bacterial Vitamin K Epoxide Reductase in the Biofilm-Forming Actinobacterium Actinomyces oris
journal, March 2017

  • Luong, Truc Thanh; Reardon-Robinson, Melissa E.; Siegel, Sara D.
  • Journal of Bacteriology, Vol. 199, Issue 10
  • DOI: 10.1128/JB.00817-16

Crystal structure of the DsbA protein required for disulphide bond formation in vivo
journal, September 1993

  • Martin, Jennifer L.; Bardwell, James C. A.; Kuriyan, John
  • Nature, Vol. 365, Issue 6445
  • DOI: 10.1038/365464a0

The CCP4 suite programs for protein crystallography
journal, September 1994


MOLREP an Automated Program for Molecular Replacement
journal, December 1997


Pi sampling: a methodical and flexible approach to initial macromolecular crystallization screening
journal, April 2011

  • Gorrec, Fabrice; Palmer, Colin M.; Lebon, Guillaume
  • Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 5
  • DOI: 10.1107/S0907444911008754

Functional Analysis of Paralogous Thiol-disulfide Oxidoreductases in Bacillus subtilis
journal, August 1999

  • Bolhuis, Albert; Venema, Gerard; Quax, Wim J.
  • Journal of Biological Chemistry, Vol. 274, Issue 35
  • DOI: 10.1074/jbc.274.35.24531

Assembly of pili on the surface of Corynebacterium diphtheriae: Corynebacterium diphtheriae pili
journal, November 2003