Pili contribute to biofilm formation in vitro in Mycobacterium tuberculosis

Saiyur Ramsugit, Sinenhlanhla Guma, Balakrishna Pillay, Paras Jain, Michelle H. Larsen, Siva Danaviah, Manormoney Pillay

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Organized bacterial communities, or biofilms, provide an important reservoir for persistent cells that are inaccessible or tolerant to antibiotics. Curli pili are cell-surface structures produced by certain bacteria and have been implicated in biofilm formation in these species. In order to determine whether these structures, which were suggested to be encoded by the Rv3312A (mtp) gene, have a similar role in Mycobacterium tuberculosis, we generated a Δmtp mutant and a mtp-complemented strain of a clinical isolate of M. tuberculosis and analyzed these strains for their ability to produce pili in comparison to the wild-type strain. Phenotypic analysis by transmission electron microscopy proved the essentiality of mtp for piliation in M. tuberculosis. We then compared biofilm formation of the derived strains in detergent-free Sauton's media. Biofilm mass was quantified spectrophotometrically using crystal violet. Furthermore, we examined mtp gene expression by quantitative real-time PCR in wild-type cells grown under biofilm versus planktonic growth conditions. We found a 68.4 % reduction in biofilm mass in the mutant compared to the wild-type strain (P = 0.002). Complementation of the mutant resulted in a restoration of the wild-type biofilm phenotype (P = 0.022). We, however, found no significant difference between mtp expression in cells of the biofilm to those growing planktonically. Our findings highlight a crucial, but non-specific, role of pili in the biofilm lifestyle of M. tuberculosis and indicate that they may represent an important target for the development of therapeutics to attenuate biofilm formation, thereby potentially reducing persistence.

Original languageEnglish (US)
Pages (from-to)725-735
Number of pages11
JournalAntonie van Leeuwenhoek, International Journal of General and Molecular Microbiology
Volume104
Issue number5
DOIs
StatePublished - Nov 2013

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Biofilms
Mycobacterium tuberculosis
In Vitro Techniques
Gentian Violet
Aptitude
Transmission Electron Microscopy
Detergents
Life Style
Real-Time Polymerase Chain Reaction
Anti-Bacterial Agents
Bacteria
Phenotype
Gene Expression

Keywords

  • Adhesin
  • Complementation
  • Curli pili
  • Gene deletion
  • Mycobacterium tuberculosis
  • Persistence

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Pili contribute to biofilm formation in vitro in Mycobacterium tuberculosis. / Ramsugit, Saiyur; Guma, Sinenhlanhla; Pillay, Balakrishna; Jain, Paras; Larsen, Michelle H.; Danaviah, Siva; Pillay, Manormoney.

In: Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology, Vol. 104, No. 5, 11.2013, p. 725-735.

Research output: Contribution to journalArticle

Ramsugit, Saiyur ; Guma, Sinenhlanhla ; Pillay, Balakrishna ; Jain, Paras ; Larsen, Michelle H. ; Danaviah, Siva ; Pillay, Manormoney. / Pili contribute to biofilm formation in vitro in Mycobacterium tuberculosis. In: Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology. 2013 ; Vol. 104, No. 5. pp. 725-735.
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