Genome-wide dissection of the quorum sensing signalling pathway in Trypanosoma brucei

Binny M Mony, Paula MacGregor, Alasdair Ivens, Federico Rojas, Andrew Cowton, Julie Young, David Horn, Keith Matthews

    Research output: Contribution to journalLetter

    93 Citations (Scopus)

    Abstract

    The protozoan parasites Trypanosoma brucei spp. cause important human and livestock diseases in sub-Saharan Africa. In mammalian blood, two developmental forms of the parasite exist: proliferative 'slender' forms and arrested 'stumpy' forms that are responsible for transmission to tsetse flies. The slender to stumpy differentiation is a density-dependent response that resembles quorum sensing in microbial systems and is crucial for the parasite life cycle, ensuring both infection chronicity and disease transmission. This response is triggered by an elusive 'stumpy induction factor' (SIF) whose intracellular signalling pathway is also uncharacterized. Laboratory-adapted (monomorphic) trypanosome strains respond inefficiently to SIF but can generate forms with stumpy characteristics when exposed to cell-permeable cAMP and AMP analogues. Exploiting this, we have used a genome-wide RNA interference library screen to identify the signalling components driving stumpy formation. In separate screens, monomorphic parasites were exposed to 8-(4-chlorophenylthio)-cAMP (pCPT-cAMP) or 8-pCPT-2'-O-methyl-5'-AMP to select cells that were unresponsive to these signals and hence remained proliferative. Genome-wide Ion Torrent based RNAi target sequencing identified cohorts of genes implicated in each step of the signalling pathway, from purine metabolism, through signal transducers (kinases, phosphatases) to gene expression regulators. Genes at each step were independently validated in cells naturally capable of stumpy formation, confirming their role in density sensing in vivo. The putative RNA-binding protein, RBP7, was required for normal quorum sensing and promoted cell-cycle arrest and transmission competence when overexpressed. This study reveals that quorum sensing signalling in trypanosomes shares similarities to fundamental quiescence pathways in eukaryotic cells, its components providing targets for quorum-sensing interference-based therapeutics.
    Original languageEnglish
    Pages (from-to)681-685
    Number of pages5
    JournalNature
    Volume505
    Issue number7485
    DOIs
    Publication statusPublished - 30 Jan 2014

    Fingerprint

    Quorum Sensing
    Trypanosoma brucei brucei
    Dissection
    Parasites
    Genome
    Trypanosomiasis
    Adenosine Monophosphate
    RNA Interference
    Tsetse Flies
    RNA-Binding Proteins
    Africa South of the Sahara
    Livestock
    Eukaryotic Cells
    Cellular Structures
    Cell Cycle Checkpoints
    Life Cycle Stages
    Transducers
    Phosphoric Monoester Hydrolases
    Mental Competency
    Genes

    Cite this

    Mony, B. M., MacGregor, P., Ivens, A., Rojas, F., Cowton, A., Young, J., ... Matthews, K. (2014). Genome-wide dissection of the quorum sensing signalling pathway in Trypanosoma brucei. Nature, 505(7485), 681-685. https://doi.org/10.1038/nature12864
    Mony, Binny M ; MacGregor, Paula ; Ivens, Alasdair ; Rojas, Federico ; Cowton, Andrew ; Young, Julie ; Horn, David ; Matthews, Keith. / Genome-wide dissection of the quorum sensing signalling pathway in Trypanosoma brucei. In: Nature. 2014 ; Vol. 505, No. 7485. pp. 681-685.
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    abstract = "The protozoan parasites Trypanosoma brucei spp. cause important human and livestock diseases in sub-Saharan Africa. In mammalian blood, two developmental forms of the parasite exist: proliferative 'slender' forms and arrested 'stumpy' forms that are responsible for transmission to tsetse flies. The slender to stumpy differentiation is a density-dependent response that resembles quorum sensing in microbial systems and is crucial for the parasite life cycle, ensuring both infection chronicity and disease transmission. This response is triggered by an elusive 'stumpy induction factor' (SIF) whose intracellular signalling pathway is also uncharacterized. Laboratory-adapted (monomorphic) trypanosome strains respond inefficiently to SIF but can generate forms with stumpy characteristics when exposed to cell-permeable cAMP and AMP analogues. Exploiting this, we have used a genome-wide RNA interference library screen to identify the signalling components driving stumpy formation. In separate screens, monomorphic parasites were exposed to 8-(4-chlorophenylthio)-cAMP (pCPT-cAMP) or 8-pCPT-2'-O-methyl-5'-AMP to select cells that were unresponsive to these signals and hence remained proliferative. Genome-wide Ion Torrent based RNAi target sequencing identified cohorts of genes implicated in each step of the signalling pathway, from purine metabolism, through signal transducers (kinases, phosphatases) to gene expression regulators. Genes at each step were independently validated in cells naturally capable of stumpy formation, confirming their role in density sensing in vivo. The putative RNA-binding protein, RBP7, was required for normal quorum sensing and promoted cell-cycle arrest and transmission competence when overexpressed. This study reveals that quorum sensing signalling in trypanosomes shares similarities to fundamental quiescence pathways in eukaryotic cells, its components providing targets for quorum-sensing interference-based therapeutics.",
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    Mony, BM, MacGregor, P, Ivens, A, Rojas, F, Cowton, A, Young, J, Horn, D & Matthews, K 2014, 'Genome-wide dissection of the quorum sensing signalling pathway in Trypanosoma brucei', Nature, vol. 505, no. 7485, pp. 681-685. https://doi.org/10.1038/nature12864

    Genome-wide dissection of the quorum sensing signalling pathway in Trypanosoma brucei. / Mony, Binny M; MacGregor, Paula; Ivens, Alasdair; Rojas, Federico; Cowton, Andrew; Young, Julie; Horn, David; Matthews, Keith.

    In: Nature, Vol. 505, No. 7485, 30.01.2014, p. 681-685.

    Research output: Contribution to journalLetter

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    AU - Horn, David

    AU - Matthews, Keith

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    Mony BM, MacGregor P, Ivens A, Rojas F, Cowton A, Young J et al. Genome-wide dissection of the quorum sensing signalling pathway in Trypanosoma brucei. Nature. 2014 Jan 30;505(7485):681-685. https://doi.org/10.1038/nature12864