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Volume 168, Issue 4

Cg1246, a new player in mycolic acid biosynthesis in Free

Abstract

Mycolic acids are key components of the complex cell envelope of . These fatty acids, conjugated to trehalose or to arabinogalactan form the backbone of the mycomembrane. While mycolic acids are essential to the survival of some species, such as , their absence is not lethal for which has been extensively used as a model to depict their biosynthesis. Mycolic acids are first synthesized on the cytoplasmic side of the inner membrane and transferred onto trehalose to give trehalose monomycolate (TMM). TMM is subsequently transported to the periplasm by dedicated transporters and used by mycoloyltransferase enzymes to synthesize all the other mycolate-containing compounds. Using a random transposition mutagenesis, we recently identified a new uncharacterized protein (Cg1246) involved in mycolic acid metabolism. Cg1246 belongs to the DUF402 protein family that contains some previously characterized nucleoside phosphatases. In this study, we performed a functional and structural characterization of Cg1246. We showed that absence of the protein led to a significant reduction in the pool of TMM in , resulting in a decrease in all other mycolate-containing compounds. We found that, , Cg1246 has phosphatase activity on organic pyrophosphate substrates but is most likely not a nucleoside phosphatase. Using a computational approach, we identified important residues for phosphatase activity and constructed the corresponding variants in . Surprisingly complementation with these non-functional proteins fully restored the defect in TMM of the Δ mutant strain, suggesting that , the phosphatase activity is not involved in mycolic acid biosynthesis.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): Corynebacterium glutamicum , lipid biosynthesis , mycolic acid , mycomembrane and phosphatase
Funding
This study was supported by the:
  • École Polytechnique, Université Paris-Saclay
    • Principle Award Recipient: Céliade Sousa-d'Auria
  • Université Paris-Saclay
    • Principle Award Recipient: Céliade Sousa-d'Auria
  • Centre National de la Recherche Scientifique
    • Principle Award Recipient: Céliade Sousa-d'Auria
© 2022 The Authors
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2022-04-08
2024-04-18
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References

  1. Daffé M, Marrakchi H. Unraveling the structure of the mycobacterial envelope. Microbiol Spectr 2019; 7: [View Article] [PubMed]
     [Google Scholar]
  2. Houssin C, de Sousa d’Auria C, Constantinesco F, Dietrich C, Labarre C et al. Architecture and biogenesis of the cell envelope of Corynebacterium glutamicum. In Inui M, Toyoda K. eds Corynebacterium Glutamicum: Biology and Biotechnology Cham: Springer International Publishing; 2020 pp 25–60
     [Google Scholar]
  3. Zuber B, Chami M, Houssin C, Dubochet J, Griffiths G et al. Direct visualization of the outer membrane of mycobacteria and corynebacteria in their native state. J Bacteriol 2008; 190:5672–5680 [View Article] [PubMed]
     [Google Scholar]
  4. Hoffmann C, Leis A, Niederweis M, Plitzko JM, Engelhardt H. Disclosure of the mycobacterial outer membrane: cryo-electron tomography and vitreous sections reveal the lipid bilayer structure. Proc Natl Acad Sci U S A 2008; 105:3963–3967 [View Article] [PubMed]
     [Google Scholar]
  5. Marrakchi H, Lanéelle M-A, Daffé M. Mycolic acids: structures, biosynthesis, and beyond. Chem Biol 2014; 21:67–85 [View Article] [PubMed]
     [Google Scholar]
  6. Quémard A. New insights into the mycolate-containing compound biosynthesis and transport in mycobacteria. Trends Microbiol 2016; 24:725–738 [View Article] [PubMed]
     [Google Scholar]
  7. Lanéelle MA, Tropis M, Daffé M. Current knowledge on mycolic acids in Corynebacterium glutamicum and their relevance for biotechnological processes. Appl Microbiol Biotechnol 2013; 97:9923–9930 [View Article] [PubMed]
     [Google Scholar]
  8. Huc E, Meniche X, Benz R, Bayan N, Ghazi A et al. O-mycoloylated proteins from Corynebacterium: An unprecedented post-translational modification in bacteria. J Biol Chem 2010; 285:21908–21912 [View Article] [PubMed]
     [Google Scholar]
  9. Huc E, de Sousa-D’Auria C, de la Sierra-Gallay IL, Salmeron C, van Tilbeurgh H et al. Identification of a mycoloyl transferase selectively involved in o-acylation of polypeptides in Corynebacteriales. J Bacteriol 2013; 195:4121–4128 [View Article] [PubMed]
     [Google Scholar]
  10. Issa H, Huc-Claustre E, Reddad T, Bonadé Bottino N, Tropis M et al. Click-chemistry approach to study mycoloylated proteins: Evidence for PorB and PorC porins mycoloylation in Corynebacterium glutamicum. PLoS One 2017; 12:e0171955 [View Article] [PubMed]
     [Google Scholar]
  11. PaweŁczyk J, Kremer L. Hatfull GF, Jacobs Jr. WR, editors The Molecular Genetics of Mycolic Acid Biosynthesis. Microbiol Spectr 2014; 2:MGM2–0003 [View Article] [PubMed]
     [Google Scholar]
  12. Radmacher E, Alderwick LJ, Besra GS, Brown AK, Gibson KJC et al. Two functional FAS-I type fatty acid synthases in Corynebacterium glutamicum. Microbiology (Reading) 2005; 151:2421–2427 [View Article] [PubMed]
     [Google Scholar]
  13. Portevin D, de Sousa-D’Auria C, Montrozier H, Houssin C, Stella A et al. The acyl-AMP ligase FadD32 and AccD4-containing acyl-CoA carboxylase are required for the synthesis of mycolic acids and essential for mycobacterial growth: identification of the carboxylation product and determination of the acyl-CoA carboxylase componen. J Biol Chem 2005; 280:8862–8874 [View Article] [PubMed]
     [Google Scholar]
  14. Portevin D, De Sousa-D’Auria C, Houssin C, Grimaldi C, Chami M et al. A polyketide synthase catalyzes the last condensation step of mycolic acid biosynthesis in mycobacteria and related organisms. Proc Natl Acad Sci U S A 2004; 101:314–319 [View Article] [PubMed]
     [Google Scholar]
  15. Gande R, Gibson KJC, Brown AK, Krumbach K, Dover LG et al. Acyl-CoA carboxylases (accD2 and accD3), together with a unique polyketide synthase (Cg-pks), are key to mycolic acid biosynthesis in Corynebacterianeae such as Corynebacterium glutamicum and Mycobacterium tuberculosis. J Biol Chem 2004; 279:44847–44857 [View Article] [PubMed]
     [Google Scholar]
  16. Gavalda S, Bardou F, Laval F, Bon C, Malaga W et al. The polyketide synthase Pks13 catalyzes a novel mechanism of lipid transfer in mycobacteria. Chem Biol 2014; 21:1660–1669 [View Article] [PubMed]
     [Google Scholar]
  17. Lea-Smith DJ, Pyke JS, Tull D, McConville MJ, Coppel RL et al. The reductase that catalyzes mycolic motif synthesis is required for efficient attachment of mycolic acids to arabinogalactan. J Biol Chem 2007; 282:11000–11008 [View Article] [PubMed]
     [Google Scholar]
  18. Varela C, Rittmann D, Singh A, Krumbach K, Bhatt K et al. MmpL genes are associated with mycolic acid metabolism in mycobacteria and corynebacteria. Chem Biol 2012; 19:498–506 [View Article] [PubMed]
     [Google Scholar]
  19. Yamaryo-Botte Y, Rainczuk AK, Lea-Smith DJ, Brammananth R, van der Peet PL et al. Acetylation of trehalose mycolates is required for efficient MmpL-mediated membrane transport in Corynebacterineae. ACS Chem Biol 2015; 10:734–746 [View Article] [PubMed]
     [Google Scholar]
  20. Rainczuk AK, Klatt S, Yamaryo-Botté Y, Brammananth R, McConville MJ et al. Mtrp, a putative methyltransferase in corynebacteria, is required for optimal membrane transport of trehalose mycolates. J Biol Chem 2020; 295:6108–6119 [View Article] [PubMed]
     [Google Scholar]
  21. Cashmore TJ, Klatt S, Brammananth R, Rainczuk AK, Crellin PK et al. MmpA, a conserved membrane protein required for efficient surface transport of trehalose lipids in Corynebacterineae. Biomolecules 2021; 11:13417–13423 [View Article] [PubMed]
     [Google Scholar]
  22. Dautin N, de Sousa-d’Auria C, Constantinesco-Becker F, Labarre C, Oberto J et al. Mycoloyltransferases: A large and major family of enzymes shaping the cell envelope of Corynebacteriales. Biochim Biophys Acta Gen Subj 2017; 1861:3581–3592 [View Article] [PubMed]
     [Google Scholar]
  23. De Sousa-D’Auria C, Kacem R, Puech V, Tropis M, Leblon G et al. New insights into the biogenesis of the cell envelope of corynebacteria: identification and functional characterization of five new mycoloyltransferase genes in Corynebacterium glutamicum. FEMS Microbiol Lett 2003; 224:35–44 [View Article] [PubMed]
     [Google Scholar]
  24. Brand S, Niehaus K, Pühler A, Kalinowski J. Identification and functional analysis of six mycolyltransferase genes of Corynebacterium glutamicum ATCC 13032: the genes cop1, cmt1, and cmt2 can replace each other in the synthesis of trehalose dicorynomycolate, a component of the mycolic acid l. Arch Microbiol 2003; 180:33–44 [View Article] [PubMed]
     [Google Scholar]
  25. Nickel J, Irzik K, van Ooyen J, Eggeling L. The TetR-type transcriptional regulator FasR of Corynebacterium glutamicum controls genes of lipid synthesis during growth on acetate. Mol Microbiol 2010; 78:253–265 [View Article] [PubMed]
     [Google Scholar]
  26. Irzik K, van Ooyen J, Gätgens J, Krumbach K, Bott M et al. Acyl-CoA sensing by FasR to adjust fatty acid synthesis in Corynebacterium glutamicum. J Biotechnol 2014; 192 Pt A:96–101 [View Article]
     [Google Scholar]
  27. Lee DS, Kim Y, Lee HS. The whcD gene of Corynebacterium glutamicum plays roles in cell division and envelope formation. Microbiol (United Kingdom) 2017; 163:131–143 [View Article]
     [Google Scholar]
  28. Lee JH, Jeong H, Kim Y, Lee HS. Corynebacterium glutamicum whiA plays roles in cell division, cell envelope formation, and general cell physiology. Antonie van Leeuwenhoek, Int J Gen Mol Microbiol 2020; 113:629–641 [View Article]
     [Google Scholar]
  29. Toyoda K, Inui M. Extracytoplasmic function sigma factor σD confers resistance to environmental stress by enhancing mycolate synthesis and modifying peptidoglycan structures in Corynebacterium glutamicum. Mol Microbiol 2018; 107:312–329 [View Article]
     [Google Scholar]
  30. Taniguchi H, Busche T, Patschkowski T, Niehaus K, Pátek M et al. Physiological roles of sigma factor SigD in Corynebacterium glutamicum. BMC Microbiol 2017; 17:158 [View Article]
     [Google Scholar]
  31. Alsayed SSR, Beh CC, Foster NR, Payne AD, Yu Y et al. Kinase targets for mycolic acid biosynthesis in Mycobacterium tuberculosis. Curr Mol Pharmacol 2019; 12:27–49 [View Article] [PubMed]
     [Google Scholar]
  32. Le N-H, Locard-Paulet M, Stella A, Tomas N, Molle V et al. The protein kinase PknB negatively regulates biosynthesis and trafficking of mycolic acids in mycobacteria. J Lipid Res 2020; 61:1180–1191 [View Article] [PubMed]
     [Google Scholar]
  33. Meniche X, Labarre C, de Sousa-d’Auria C, Huc E, Laval F et al. Identification of a stress-induced factor of Corynebacterineae that is involved in the regulation of the outer membrane lipid composition. J Bacteriol 2009; 191:7323–7332 [View Article] [PubMed]
     [Google Scholar]
  34. de Sousa-d’Auria C, Constantinesco-Becker F, Constant P, Tropis M, Houssin C. Genome-wide identification of novel genes involved in Corynebacteriales cell envelope biogenesis using Corynebacterium glutamicum as a model. PLoS One 2020; 15:e0240497 [View Article] [PubMed]
     [Google Scholar]
  35. Woodyer RD, Shao Z, Thomas PM, Kelleher NL, Blodgett JAV et al. Heterologous production of fosfomycin and identification of the minimal biosynthetic gene cluster. Chem Biol 2006; 13:1171–1182 [View Article] [PubMed]
     [Google Scholar]
  36. Sato S, Miyanaga A, Kim S-Y, Kuzuyama T, Kudo F et al. Biochemical and structural analysis of FomD that catalyzes the hydrolysis of cytidylyl (S)-2-hydroxypropylphosphonate in fosfomycin biosynthesis. Biochemistry 2018; 57:4858–4866 [View Article] [PubMed]
     [Google Scholar]
  37. Imae K, Saito Y, Kizaki H, Ryuno H, Mao H et al. Novel nucleoside diphosphatase contributes to Staphylococcus aureus virulence. J Biol Chem 2016; 291:18608–18619 [View Article] [PubMed]
     [Google Scholar]
  38. Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual, 2nd ed. New York: Cold Spring Harbor Laboratory, Cold Spring Harbor; 1989
     [Google Scholar]
  39. Ausubel F, Brent R, Kingston R, Moore D, Seidman J et al. Current Protocols in Molecular Biology Curr Protoc Mol Biol New York: Wiley Interscience; 1987
     [Google Scholar]
  40. Schäfer A, Tauch A, Jäger W, Kalinowski J, Thierbach G et al. Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 1994; 145:69–73 [View Article] [PubMed]
     [Google Scholar]
  41. Peyret JL, Bayan N, Joliff G, Gulik-Krzywicki T, Mathieu L et al. Characterization of the cspB gene encoding PS2, an ordered surface-layer protein in Corynebacterium glutamicum. Mol Microbiol 1993; 9:97–109 [View Article] [PubMed]
     [Google Scholar]
  42. Song Y, DiMaio F, Wang RY-R, Kim D, Miles C et al. High-resolution comparative modeling with RosettaCM. Structure 2013; 21:1735–1742 [View Article] [PubMed]
     [Google Scholar]
  43. Dusch N, Pühler A, Kalinowski J. Expression of the Corynebacterium glutamicum panD gene encoding L-aspartate-alpha-decarboxylase leads to pantothenate overproduction in Escherichia coli. Appl Environ Microbiol 1999; 65:1530–1539 [View Article] [PubMed]
     [Google Scholar]
  44. Reimer LC, Spura J, Schmidt-Hohagen K, Schomburg D. High-throughput screening of a Corynebacterium glutamicum mutant library on genomic and metabolic level. PLoS One 2014; 9:e86799 [View Article] [PubMed]
     [Google Scholar]
  45. Silberbach M, Schäfer M, Hüser AT, Kalinowski J, Pühler A et al. Adaptation of Corynebacterium glutamicum to ammonium limitation: a global analysis using transcriptome and proteome techniques. Appl Environ Microbiol 2005; 71:2391–2402 [View Article] [PubMed]
     [Google Scholar]
  46. Rey DA, Nentwich SS, Koch DJ, Rückert C, Pühler A et al. The McbR repressor modulated by the effector substance S- adenosylhomocysteine controls directly the transcription of a regulon involved in sulphur metabolism of Corynebacterium glutamicum ATCC 13032. Mol Microbiol 2005; 56:871–887 [View Article] [PubMed]
     [Google Scholar]
  47. Jeffery CJ. Moonlighting proteins: Old proteins learning new tricks. Trends Genet 2003; 19:415–417 [View Article] [PubMed]
     [Google Scholar]
  48. Jeffery CJ. What is Protein Moonlighting and Why is it Important?. In Henderson B. eds Moonlighting Proteins: Novel Virulence Factors in Bacterial Infections 2017 pp 1–19 [View Article]
     [Google Scholar]
  49. Mani M, Chen C, Amblee V, Liu H, Mathur T et al. MoonProt: a database for proteins that are known to moonlight. Nucleic Acids Res 2015; 43:D277–82 [View Article] [PubMed]
     [Google Scholar]
  50. Banerjee S, Nandyala AK, Raviprasad P, Ahmed N, Hasnain SE. Iron-dependent RNA-binding activity of Mycobacterium tuberculosis aconitase. J Bacteriol 2007; 189:4046–4052 [View Article] [PubMed]
     [Google Scholar]
  51. Commichau FM, Stülke J. Trigger enzymes: bifunctional proteins active in metabolism and in controlling gene expression. Mol Microbiol 2008; 67:692–702 [View Article] [PubMed]
     [Google Scholar]
  52. Ashkenazy H, Abadi S, Martz E, Chay O, Mayrose I et al. ConSurf 2016: an improved methodology to estimate and visualize evolutionary conservation in macromolecules. Nucleic Acids Res 2016; 44:W344–50 [View Article] [PubMed]
     [Google Scholar]
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Cg1246, a new player in mycolic acid biosynthesis in Corynebacterium glutamicum
Microbiology 168, 001171 (2022); https://doi.org/10.1099/mic.0.001171
/content/journal/micro/10.1099/mic.0.001171
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