Cooperative retraction of bundled type IV pili enables nanonewton force generation

Nicolas Biais, Benoît Ladoux, Dustin Higashi, Magdalene So, Michael Sheetz

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

The causative agent of gonorrhea, Neisseria gonorrhoeae, bears retractable filamentous appendages called type IV pili (Tfp). Tfp are used by many pathogenic and nonpathogenic bacteria to carry out a number of vital functions, including DNA uptake, twitching motility (crawling over surfaces), and attachment to host cells. In N. gonorrhoeae, Tfp binding to epithelial cells and the mechanical forces associated with this binding stimulate signaling cascades and gene expression that enhance infection. Retraction of a single Tfp filament generates forces of 50-100 piconewtons, but nothing is known, thus far, on the retraction force ability of multiple Tfp filaments, even though each bacterium expresses multiple Tfp and multiple bacteria interact during infection. We designed a micropillar assay system to measure Tfp retraction forces. This system consists of an array of force sensors made of elastic pillars that allow quantification of retraction forces from adherent N. gonorrhoeae bacteria. Electron microscopy and fluorescence microscopy were used in combination with this novel assay to assess the structures of Tfp. We show that Tfp can form bundles, which contain up to 8-10 Tfp filaments, that act as coordinated retractable units with forces up to 10 times greater than single filament retraction forces. Furthermore, single filament retraction forces are transient, whereas bundled filaments produce retraction forces that can be sustained. Alterations of noncovalent protein-protein interactions between Tfp can inhibit both bundle formation and high-amplitude retraction forces. Retraction forces build over time through the recruitment and bundling of multiple Tfp that pull cooperatively to generate forces in the nanonewton range. We propose that Tfp retraction can be synchronized through bundling, that Tfp bundle retraction can generate forces in the nanonewton range in vivo, and that such high forces could affect infection.

Original languageEnglish (US)
Article numbere87
Pages (from-to)907-913
Number of pages7
JournalPLoS Biology
Volume6
Issue number4
DOIs
StatePublished - Apr 1 2008
Externally publishedYes

Fingerprint

fimbriae
cooperatives
Neisseria gonorrhoeae
Bacteria
Assays
Infection
Gonorrhea
Fluorescence microscopy
Fluorescence Microscopy
Gene expression
Electron microscopy
Electron Microscopy
Proteins
Epithelial Cells
Gene Expression
bacteria
DNA
Sensors
infection
protein-protein interactions

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

Cooperative retraction of bundled type IV pili enables nanonewton force generation. / Biais, Nicolas; Ladoux, Benoît; Higashi, Dustin; So, Magdalene; Sheetz, Michael.

In: PLoS Biology, Vol. 6, No. 4, e87, 01.04.2008, p. 907-913.

Research output: Contribution to journalArticle

Biais, Nicolas ; Ladoux, Benoît ; Higashi, Dustin ; So, Magdalene ; Sheetz, Michael. / Cooperative retraction of bundled type IV pili enables nanonewton force generation. In: PLoS Biology. 2008 ; Vol. 6, No. 4. pp. 907-913.
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