1887

Abstract

Iron-sulphur (FeS) clusters are versatile cofactors required for a range of biological processes within cells. Due to the reactive nature of the constituent molecules, assembly and delivery of these cofactors requires a multi-protein machinery . In prokaryotes, SufT homologues are proposed to function in the maturation and transfer of FeS clusters to apo-proteins. This study used targeted gene deletion to investigate the role of SufT in the physiology of mycobacteria, using as a model organism. Deletion of the gene in had no impact on growth under standard culture conditions and did not significantly alter activity of the FeS cluster dependent enzymes succinate dehydrogenase (SDH) and aconitase (ACN). Furthermore, the Δ mutant was no more sensitive than the wild-type strain to the redox cycler 2,3-dimethoxy-1,4-naphthoquinone (DMNQ), or the anti-tuberculosis drugs isoniazid, clofazimine or rifampicin. In contrast, the Δ mutant displayed a growth defect under iron limiting conditions, and an increased requirement for iron during biofilm formation. This data suggests that SufT is an accessory factor in FeS cluster biogenesis in mycobacteria which is required under conditions of iron limitation.

Funding
This study was supported by the:
  • South African Medical Research Council (Award CTBR)
    • Principle Award Recipient: Not Applicable
  • National Research Foundation (ZA) (Award 81781)
    • Principle Award Recipient: Not Applicable
  • National Research Foundation (ZA) (Award 91424)
    • Principle Award Recipient: Monique J. Williams
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000881
2019-12-20
2024-11-02
Loading full text...

Full text loading...

/deliver/fulltext/micro/166/3/296.html?itemId=/content/journal/micro/10.1099/mic.0.000881&mimeType=html&fmt=ahah

References

  1. WHO 2018; Global tuberculosis report 2018. World health organisation. https://www.who.int/tb/publications/global_report/en/ 24 June 2019
  2. Mao C, Shukla M, Larrouy-Maumus G, Dix FL, Kelley LA et al. Functional assignment of Mycobacterium tuberculosis proteome revealed by genome-scale fold-recognition. Tuberculosis 2013; 93:40–46 [View Article][PubMed]
    [Google Scholar]
  3. Cole ST, Brosch R, Parkhill J, Garnier T, Churcher C et al. Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature 1998; 393:537–544 [View Article][PubMed]
    [Google Scholar]
  4. Huet G, Daffé M, Saves I. Identification of the Mycobacterium tuberculosis SUF machinery as the exclusive mycobacterial system of [Fe-S] cluster assembly: evidence for its implication in the pathogen's survival. J Bacteriol 2005; 187:6137–6146 [View Article][PubMed]
    [Google Scholar]
  5. Sassetti CM, Boyd DH, Rubin EJ. Genes required for mycobacterial growth defined by high density mutagenesis. Mol Microbiol 2003; 48:77–84 [View Article][PubMed]
    [Google Scholar]
  6. Griffin JE, Gawronski JD, Dejesus MA, Ioerger TR, Akerley BJ et al. High-Resolution phenotypic profiling defines genes essential for mycobacterial growth and cholesterol catabolism. PLoS Pathog 2011; 7:e1002251 [View Article][PubMed]
    [Google Scholar]
  7. Cole ST, Eiglmeier K, Parkhill J, James KD, Thomson NR et al. Massive gene decay in the leprosy Bacillus. Nature 2001; 409:1007–1011 [View Article][PubMed]
    [Google Scholar]
  8. Willemse D, Weber B, Masino L, Warren RM, Adinolfi S et al. Rv1460, a SufR homologue, is a repressor of the suf operon in Mycobacterium tuberculosis . PLoS One 2018; 13:e0200145 [View Article][PubMed]
    [Google Scholar]
  9. Wollers S, Layer G, Garcia-Serres R, Signor L, Clemancey M et al. Iron-Sulfur (Fe-S) cluster assembly: the SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor. J Biol Chem 2010; 285:23331–23341 [View Article][PubMed]
    [Google Scholar]
  10. Hirabayashi K, Yuda E, Tanaka N, Katayama S, Iwasaki K et al. Functional dynamics revealed by the structure of the SufBCD complex, a novel ATP-binding cassette (ABC) protein that serves as a scaffold for iron-sulfur cluster biogenesis. J Biol Chem 2015; 290:29717–29731 [View Article][PubMed]
    [Google Scholar]
  11. Rybniker J, Pojer F, Marienhagen J, Kolly GS, Chen JM et al. The cysteine desulfurase IscS of Mycobacterium tuberculosis is involved in iron–sulfur cluster biogenesis and oxidative stress defence. Biochem J 2014; 459:467–478 [View Article]
    [Google Scholar]
  12. Mashruwala AA, Bhatt S, Poudel S, Boyd ES, Boyd JM. The DUF59 Containing Protein SufT Is Involved in the Maturation of Iron-Sulfur (FeS) Proteins during Conditions of High FeS Cofactor Demand in Staphylococcus aureus . PLoS Genet 2016; 12:e1006233 [View Article][PubMed]
    [Google Scholar]
  13. Mashruwala AA, Pang YY, Rosario-Cruz Z, Chahal HK, Benson MA et al. Nfu facilitates the maturation of iron-sulfur proteins and participates in virulence in Staphylococcus aureus . Mol Microbiol 2015; 95:383–409 [View Article][PubMed]
    [Google Scholar]
  14. Mashruwala AA, Roberts CA, Bhatt S, May KL, Carroll RK et al. Staphylococcus aureus SufT: An essential iron-sulfur cluster assembly factor in cells experiencing a high-demand for lipoic acid. Mol Microbiol
    [Google Scholar]
  15. Sasaki S, Minamisawa K, Mitsui H. A Sinorhizobium meliloti RpoH-Regulated Gene Is Involved in Iron-Sulfur Protein Metabolism and Effective Plant Symbiosis under Intrinsic Iron Limitation. J Bacteriol 2016; 198:2297–2306 [View Article][PubMed]
    [Google Scholar]
  16. Parish T, Stoker NG. Use of a flexible cassette method to generate a double unmarked Mycobacterium tuberculosis tlyA plcABC mutant by gene replacement. Microbiology 2000; 146:1969–1975 [View Article][PubMed]
    [Google Scholar]
  17. Munujos P, Coll-Cantí J, González-Sastre F, Gella FJ. Assay of succinate dehydrogenase activity by a colorimetric-continuous method using iodonitrotetrazolium chloride as electron acceptor. Anal Biochem 1993; 212:506–509 [View Article][PubMed]
    [Google Scholar]
  18. Wallace RJ, Nash DR, Steele LC, Steingrube V. Susceptibility testing of slowly growing mycobacteria by a microdilution MIC method with 7H9 broth. J Clin Microbiol 1986; 24:976–981[PubMed]
    [Google Scholar]
  19. Kulka K, Hatfull G, Ojha AK. Growth of Mycobacterium tuberculosis biofilms. J Vis Exp 20123820 [View Article][PubMed]
    [Google Scholar]
  20. DeJesus MA, Gerrick ER, Xu W, Park SW, Long JE et al. Comprehensive Essentiality Analysis of the Mycobacterium tuberculosis Genome via Saturating Transposon Mutagenesis. mBio 2017; 8:e02133-16 [View Article][PubMed]
    [Google Scholar]
  21. Imlay JA. Iron-Sulphur clusters and the problem with oxygen. Mol Microbiol 2006; 59:1073–1082 [View Article]
    [Google Scholar]
  22. Jang S, Imlay JA. Hydrogen peroxide inactivates the Escherichia coli Isc iron-sulphur assembly system, and OxyR induces the Suf system to compensate. Mol Microbiol 2010; 78:1448–1467 [View Article][PubMed]
    [Google Scholar]
  23. Schnappinger D, Ehrt S, Voskuil MI, Liu Y, Mangan JA et al. Transcriptional adaptation of Mycobacterium tuberculosis within macrophages: insights into the phagosomal environment. J Exp Med 2003; 198:693–704 [View Article][PubMed]
    [Google Scholar]
  24. Wang J-Y, Burger RM, Drlica K. Role of superoxide in Catalase-Peroxidase-Mediated isoniazid action against mycobacteria. Antimicrob Agents Chemother 1998; 42:709–711 [View Article]
    [Google Scholar]
  25. Warek U, Falkinham JO. Action of clofazimine on the Mycobacterium avium complex. Res Microbiol 1996; 147:43–48 [View Article][PubMed]
    [Google Scholar]
  26. Kurthkoti K, Amin H, Marakalala MJ, Ghanny S, Subbian S et al. The Capacity of Mycobacterium tuberculosis To Survive Iron Starvation Might Enable It To Persist in Iron-Deprived Microenvironments of Human Granulomas. mBio 2017; 8:e01092-17 [View Article][PubMed]
    [Google Scholar]
  27. Ojha A, Hatfull GF. The role of iron in Mycobacterium smegmatis biofilm formation: the exochelin siderophore is essential in limiting iron conditions for biofilm formation but not for planktonic growth. Mol Microbiol 2007; 66:468–483 [View Article][PubMed]
    [Google Scholar]
  28. Reddy PV, Puri RV, Khera A, Tyagi AK. Iron storage proteins are essential for the survival and pathogenesis of Mycobacterium tuberculosis in THP-1 macrophages and the guinea pig model of infection. J Bacteriol 2012; 194:567–575 [View Article][PubMed]
    [Google Scholar]
  29. Reddy PV, Puri RV, Chauhan P, Kar R, Rohilla A et al. Disruption of mycobactin biosynthesis leads to attenuation of Mycobacterium tuberculosis for growth and virulence. J Infect Dis 2013; 208:1255–1265 [View Article][PubMed]
    [Google Scholar]
  30. Golden CA, Kochan I, Spriggs DR. Role of mycobactin in the growth and virulence of tubercle bacilli. Infect Immun 1974; 9:34–40[PubMed]
    [Google Scholar]
  31. Pandey R, Rodriguez GM. A ferritin mutant of Mycobacterium tuberculosis is highly susceptible to killing by antibiotics and is unable to establish a chronic infection in mice. Infect Immun 2012; 80:3650–3659 [View Article][PubMed]
    [Google Scholar]
  32. Boelaert JR, Vandecasteele SJ, Appelberg R, Gordeuk VR. The effect of the host's iron status on tuberculosis. J Infect Dis 2007; 195:1745–1753 [View Article][PubMed]
    [Google Scholar]
  33. Parrow NL, Fleming RE, Minnick MF. Sequestration and scavenging of iron in infection. Infect Immun 2013; 81:3503–3514 [View Article][PubMed]
    [Google Scholar]
  34. Dragset MS, Ioerger TR, Zhang YJ, Mærk M, Ginbot Z et al. Genome-Wide phenotypic profiling identifies and Categorizes genes required for mycobacterial low iron fitness. Sci Rep 2019; 9:1–11 [View Article]
    [Google Scholar]
  35. Ojha A, Anand M, Bhatt A, Kremer L, Jacobs WR et al. Groel1: a dedicated chaperone involved in mycolic acid biosynthesis during biofilm formation in mycobacteria. Cell 2005; 123:861–873 [View Article][PubMed]
    [Google Scholar]
  36. Yang Y, Thomas J, Li Y, Vilchèze C, Derbyshire KM et al. Defining a temporal order of genetic requirements for development of mycobacterial biofilms. Mol Microbiol 2017; 105:794809 [View Article][PubMed]
    [Google Scholar]
  37. Ojha AK, Baughn AD, Sambandan D, Hsu T, Trivelli X et al. Growth of Mycobacterium tuberculosis biofilms containing free mycolic acids and harbouring drug-tolerant bacteria. Mol Microbiol 2008; 69:164–174 [View Article][PubMed]
    [Google Scholar]
  38. Ojha AK, Trivelli X, Guerardel Y, Kremer L, Hatfull GF. Enzymatic hydrolysis of trehalose dimycolate releases free mycolic acids during mycobacterial growth in biofilms. J Biol Chem 2010; 285:17380–17389 [View Article][PubMed]
    [Google Scholar]
  39. Saini V, Farhana A, Glasgow JN, Steyn AJC. Iron sulfur cluster proteins and microbial regulation: implications for understanding tuberculosis. Curr Opin Chem Biol 2012; 16:45–53 [View Article][PubMed]
    [Google Scholar]
  40. Santos TMA, Lammers MG, Zhou M, Sparks IL, Rajendran M et al. Small molecule chelators reveal that iron starvation inhibits late stages of bacterial cytokinesis. ACS Chem Biol 2018; 13:235–246 [View Article]
    [Google Scholar]
  41. Vijay S, Vinh DN, Hai HT, Ha VTN, Dung VTM et al. Influence of Stress and Antibiotic Resistance on Cell-Length Distribution in Mycobacterium tuberculosis Clinical Isolates. Front Microbiol 2017; 8: [View Article]
    [Google Scholar]
/content/journal/micro/10.1099/mic.0.000881
Loading
/content/journal/micro/10.1099/mic.0.000881
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error