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dc.contributor.authorMontero, Manuel-
dc.contributor.authorRahimpour, Mehdi-
dc.contributor.authorViale, Alejandro M.-
dc.contributor.authorAlmagro, Goizeder-
dc.contributor.authorEydallin, Gustavo-
dc.contributor.authorMuñoz Pérez, Francisco José-
dc.contributor.authorBaroja-Fernández, Edurne-
dc.contributor.authorBahaji, Abdellatif-
dc.contributor.authorPozueta Romero, Javier-
dc.date.accessioned2015-11-27T08:10:25Z-
dc.date.available2015-11-27T08:10:25Z-
dc.date.issued2014-09-04-
dc.identifierdoi: 10.1371/journal.pone.0106938-
dc.identifierissn: 1932-6203-
dc.identifier.citationPLoS ONE 9(9): e106938 (2014)-
dc.identifier.urihttp://hdl.handle.net/10261/126049-
dc.descriptionPozueta Romero, Javier et al.-
dc.description.abstractIn Escherichia coli, ppGpp is a major determinant of growth and glycogen accumulation. Levels of this signaling nucleotide are controlled by the balanced activities of the ppGpp RelA synthetase and the dual-function hydrolase/synthetase SpoT. Here we report the construction of spoT null (DspoT) mutants obtained by transducing a DspoT allele from DrelADspoT double mutants into relA+ cells. Iodine staining of randomly selected transductants cultured on a rich complex medium revealed differences in glycogen content among them. Sequence and biochemical analyses of 8 DspoT clones displaying glycogen-deficient phenotypes revealed different inactivating mutations in relA and no detectable ppGpp when cells were cultured on a rich complex medium. Remarkably, although the co-existence of DspoT with relA proficient alleles has generally been considered synthetically lethal, we found that 11 DspoT clones displaying high glycogen phenotypes possessed relA mutant alleles with non-inactivating mutations that encoded stable RelA proteins and ppGpp contents reaching 45-85% of those of wild type cells. None of the DspoT clones, however, could grow on M9-glucose minimal medium. Both Sanger sequencing of specific genes and high-throughput genome sequencing of the DspoT clones revealed that suppressor mutations were restricted to the relA locus. The overall results (a) defined in around 4 nmoles ppGpp/g dry weight the threshold cellular levels that suffice to trigger net glycogen accumulation, (b) showed that mutations in relA, but not necessarily inactivating mutations, can be selected to compensate total SpoT function(s) loss, and (c) provided useful tools for studies of the in vivo regulation of E. coli RelA ppGpp synthetase.-
dc.description.sponsorshipThis research was partially supported by the Comisión Interministerial de Ciencia y Tecnología and Fondo Europeo de Desarrollo Regional (Spain) [grant numbers BIO2010-18239 and BIO2011-29233-002-01], the Fundación Séneca [grant number 08660/P1/08] and JSPS (Japan Society for the Promotion of Science) KAKENHI Grant-in-Aid for Scientific Research (A) [grant number 22241050]. GA and GE acknowledge fellowships from the Public University of Navarra. MR acknowledges a pre-doctoral JAE fellowship from the Consejo Superior de Investigaciones Científicas. AMV is grateful to the funding of the Programa Campus Ibericus de Excelencia Internacional, Ministerio de Educación, Spain. His 2-months visit (January–March 2014) to the Institute of Agrobiotechnology, Public University of Navarra, Pamplona, Spain, was included into the Proyecto financiado por el Ministerio de Educación en el marco del Programa Campus de Excelencia Internacional.-
dc.publisherPublic Library of Science-
dc.relation.isversionofPublisher's version-
dc.rightsopenAccess-
dc.titleSystematic Production of Inactivating and Non-Inactivating Suppressor Mutations at the relA Locus That Compensate the Detrimental Effects of Complete spoT Loss and Affect Glycogen Content in Escherichia coli-
dc.typeartículo-
dc.identifier.doi10.1371/journal.pone.0106938-
dc.relation.publisherversionhttp://dx.doi.org/10.1371/journal.pone.0106938-
dc.date.updated2015-11-27T08:10:25Z-
dc.description.versionPeer Reviewed-
dc.language.rfc3066eng-
dc.rights.licensehttp://creativecommons.org/licenses/by/4.0/-
dc.contributor.funderFundación Séneca-
dc.contributor.funderMinisterio de Educación (España)-
dc.contributor.funderComisión Interministerial de Ciencia y Tecnología, CICYT (España)-
dc.contributor.funderEuropean Commission-
dc.contributor.funderConsejo Superior de Investigaciones Científicas (España)-
dc.contributor.funderUniversidad de Navarra-
dc.contributor.funderMinistry of Education, Culture, Sports, Science and Technology (Japan)-
dc.contributor.funderJapan Society for the Promotion of Science-
dc.relation.csicSí-
dc.identifier.funderhttp://dx.doi.org/10.13039/100007801es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100007273es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100000780es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100003339es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100004435es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100001700es_ES
dc.identifier.funderhttp://dx.doi.org/10.13039/501100001691es_ES
dc.identifier.pmid25188023-
dc.type.coarhttp://purl.org/coar/resource_type/c_6501es_ES
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.fulltextWith Fulltext-
item.cerifentitytypePublications-
item.openairetypeartículo-
item.grantfulltextopen-
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