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Title

ParB dynamics and the critical role of the CTD in DNA condensation unveiled by combined force-fluorescence measurements

AuthorsMadariaga-Marcos, Julene; Pastrana, Cesar L.; Fisher, Gemma LM; Dillingham, Mark Simon; Moreno-Herrero, Fernando
KeywordsB. subtilis
ParB
Spo0J
TIRF microscopy
Bacterial chromosome dynamic
Chromosome
Gene expression
Magnetic tweezers
Molecular biophysics;
Single molecule
Structural biology
Issue DateMar-2019
PublishereLife Sciences Publications
CitationeLife 8: e43812 (2019)
AbstractBacillus subtilis ParB forms multimeric networks involving non-specific DNA binding leading to DNA condensation. Previously, we found that an excess of the free C-terminal domain (CTD) of ParB impeded DNA condensation or promoted decondensation of pre-assembled networks (Fisher et al., 2017). However, interpretation of the molecular basis for this phenomenon was complicated by our inability to uncouple protein binding from DNA condensation. Here, we have combined lateral magnetic tweezers with TIRF microscopy to simultaneously control the restrictive force against condensation and to visualise ParB protein binding by fluorescence. At non-permissive forces for condensation, ParB binds non-specifically and highly dynamically to DNA. Our new approach concluded that the free CTD blocks the formation of ParB networks by heterodimerisation with full length DNA-bound ParB. This strongly supports a model in which the CTD acts as a key bridging interface between distal DNA binding loci within ParB networks.
Publisher version (URL)http://dx.doi.org/10.7554/eLife.43812.
URIhttp://hdl.handle.net/10261/179418
DOI10.7554/eLife.43812
ISSN2050-084X
E-ISSN2050-084X
Appears in Collections:(CNB) Artículos
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