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Article Dans Une Revue PLoS ONE Année : 2020

Addressing the role of centromere sites in activation of ParB proteins for partition complex assembly

Sylvain Audibert
  • Fonction : Auteur
Nicolas Tanguy-Le-Gac
Jérôme Rech
  • Fonction : Auteur
Catherine Turlan
  • Fonction : Auteur
Kerstin Bystricky
David Lane
  • Fonction : Auteur

Résumé

The ParB-parS partition complexes that bacterial replicons use to ensure their faithful inheritance also find employment in visualization of DNA loci, as less intrusive alternatives to fluorescent repressor-operator systems. The ability of ParB molecules to interact via their Nterminal domains and to bind to non-specific DNA enables expansion of the initial complex to a size both functional in partition and, via fusion to fluorescent peptides, visible by light microscopy. We have investigated whether it is possible to dispense with the need to insert parS in the genomic locus of interest, by determining whether ParB fused to proteins that bind specifically to natural DNA sequences can still assemble visible complexes. In yeast cells, coproduction of fusions of ParB to a fluorescent peptide and to a TALE protein targeting an endogenous sequence did not yield visible foci; nor did any of several variants of these components. In E.coli, coproduction of fusions of SopB (F plasmid ParB) to fluorescent peptide, and to dCas9 together with specific guide RNAs, likewise yielded no foci. The result of coproducing analogous fusions of SopB proteins with distinct binding specificities was also negative. Our observations imply that in order to assemble higher order partition complexes, ParB proteins need specific activation through binding to their cognate parS sites.
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Dates et versions

hal-02976781 , version 1 (10-12-2020)

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Citer

Sylvain Audibert, Nicolas Tanguy-Le-Gac, Jérôme Rech, Catherine Turlan, Jean-Yves Bouet, et al.. Addressing the role of centromere sites in activation of ParB proteins for partition complex assembly. PLoS ONE, 2020, 15 (5), pp.e0226472. ⟨10.1371/journal.pone.0226472⟩. ⟨hal-02976781⟩
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