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dc.contributor.authorRao, Chitong
dc.contributor.authorGuyard, Cyril
dc.contributor.authorPelaz, Carmen 
dc.contributor.authorWasserscheid, Jessica
dc.contributor.authorBondy-Denomy, Joseph
dc.contributor.authorDewar, Ken
dc.contributor.authorEnsminger, Alexander W
dc.date.accessioned2021-04-29T10:32:56Z
dc.date.available2021-04-29T10:32:56Z
dc.date.issued2016
dc.identifier.citationCell Microbiol . 2016 Oct;18(10):1319-38.es_ES
dc.identifier.urihttp://hdl.handle.net/20.500.12105/12818
dc.description.abstractClustered regularly interspaced short palindromic repeats with CRISPR-associated gene (CRISPR-Cas) systems are widely recognized as critical genome defense systems that protect microbes from external threats such as bacteriophage infection. Several isolates of the intracellular pathogen Legionella pneumophila possess multiple CRISPR-Cas systems (type I-C, type I-F and type II-B), yet the targets of these systems remain unknown. With the recent observation that at least one of these systems (II-B) plays a non-canonical role in supporting intracellular replication, the possibility remained that these systems are vestigial genome defense systems co-opted for other purposes. Our data indicate that this is not the case. Using an established plasmid transformation assay, we demonstrate that type I-C, I-F and II-B CRISPR-Cas provide protection against spacer targets. We observe efficient laboratory acquisition of new spacers under 'priming' conditions, in which initially incomplete target elimination leads to the generation of new spacers and ultimate loss of the invasive DNA. Critically, we identify the first known target of L. pneumophila CRISPR-Cas: a 30 kb episome of unknown function whose interbacterial transfer is guarded against by CRISPR-Cas. We provide evidence that the element can subvert CRISPR-Cas by mutating its targeted sequences - but that primed spacer acquisition may limit this mechanism of escape. Rather than generally impinging on bacterial fitness, this element drives a host specialization event - with improved fitness in Acanthamoeba but a reduced ability to replicate in other hosts and conditions. These observations add to a growing body of evidence that host range restriction can serve as an existential threat to L. pneumophila in the wild.es_ES
dc.description.sponsorshipThis work was supported by the University of Toronto and operating grants awarded to AWE by the Canadian Institutes of Health Research (MOP‐133406) and the Natural Sciences and Engineering Research Council of Canada (RGPIN‐2014‐03641), along with an infrastructure grant awarded to AWE from the Canada Foundation for Innovation and the Ontario Research Fund (30364). The authors have no conflict of interest to declare.es_ES
dc.language.isoenges_ES
dc.publisherWiley es_ES
dc.type.hasVersionVoRes_ES
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subject.meshClustered Regularly Interspaced Short Palindromic Repeats es_ES
dc.subject.meshAcanthamoeba castellanii es_ES
dc.subject.meshBase Sequence es_ES
dc.subject.meshConserved Sequence es_ES
dc.subject.meshEvolution, Molecular es_ES
dc.subject.meshGenes, Bacterial es_ES
dc.subject.meshHost-Pathogen Interactions es_ES
dc.subject.meshLegionella pneumophila es_ES
dc.subject.meshMicrobial Viability es_ES
dc.subject.meshSequence Analysis, DNA es_ES
dc.titleActive and adaptive Legionella CRISPR-Cas reveals a recurrent challenge to the pathogen.es_ES
dc.typejournal articlees_ES
dc.rights.licenseAtribución 4.0 Internacional*
dc.identifier.pubmedID26936325es_ES
dc.format.volume18es_ES
dc.format.number10es_ES
dc.format.page1319-38es_ES
dc.identifier.doi10.1111/cmi.12586es_ES
dc.contributor.funderUniversity of Toronto (Cánada) 
dc.contributor.funderCanadian Institutes of Health Research 
dc.contributor.funderNatural Sciences and Engineering Research Council (Canada) 
dc.contributor.funderCanada Foundation for Innovation 
dc.contributor.funderOntario Research Fund - Research Excellence (ORF-RE) 
dc.description.peerreviewedes_ES
dc.identifier.e-issn1462-5822es_ES
dc.relation.publisherversionhttps://doi.org/10.1111/cmi.12586es_ES
dc.identifier.journalCellular microbiologyes_ES
dc.repisalud.centroISCIII::Centro Nacional de Microbiologíaes_ES
dc.repisalud.institucionISCIIIes_ES
dc.rights.accessRightsopen accesses_ES


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Atribución 4.0 Internacional
Este Item está sujeto a una licencia Creative Commons: Atribución 4.0 Internacional