Addressing the role of centromere sites in activation of ParB proteins for partition complex assembly
| Autor: | Nicolas Tanguy-le-Gac, Catherine Turlan, Jean-Yves Bouet, Sylvain Audibert, David Lane, Jérôme Rech, Kerstin Bystricky |
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| Přispěvatelé: | Laboratoire de microbiologie et génétique moléculaires (LMGM), Centre de Biologie Intégrative (CBI), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Molecular biology
[SDV]Life Sciences [q-bio] medicine.disease_cause Biochemistry Cell Fusion chemistry.chemical_compound 0302 clinical medicine Plasmid CRISPR-Associated Protein 9 Protein targeting Replicon Guide RNA Materials Centromeres 0303 health sciences Multidisciplinary Symporters Chromosome Biology Organic Compounds Chemistry Escherichia coli Proteins Monosaccharides Eukaryota Cell biology Physical Sciences Medicine Plasmids Protein Binding Research Article DNA Bacterial Chromosome Structure and Function Cell Physiology Saccharomyces cerevisiae Proteins Recombinant Fusion Proteins Science Centromere Green Fluorescent Proteins Materials Science Protein domain Carbohydrates Saccharomyces cerevisiae DNA construction DNA-binding protein Chromosomes 03 medical and health sciences Bacterial Proteins Protein Domains DNA-binding proteins Escherichia coli medicine Dimers 030304 developmental biology Binding Sites Base Sequence Biology and life sciences Organic Chemistry Organisms Fungi Chemical Compounds Proteins Cell Biology Polymer Chemistry Arabinose Yeast Research and analysis methods Molecular biology techniques Oligomers Plasmid Construction 030217 neurology & neurosurgery DNA |
| Zdroj: | PLoS ONE, Vol 15, Iss 5, p e0226472 (2020) PLoS ONE PLoS ONE, Public Library of Science, 2020, 15 (5), pp.e0226472. ⟨10.1371/journal.pone.0226472⟩ |
| ISSN: | 1932-6203 |
| Popis: | International audience; 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. |
| Databáze: | OpenAIRE |
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