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dc.contributor.authorDarbari, VCen_US
dc.contributor.authorLawton, Een_US
dc.contributor.authorLu, Den_US
dc.contributor.authorBurrows, PCen_US
dc.contributor.authorWiesler, Sen_US
dc.contributor.authorJoly, Nen_US
dc.contributor.authorZhang, Nen_US
dc.contributor.authorZhang, Xen_US
dc.contributor.authorBuck, Men_US
dc.date.accessioned2016-06-20T11:29:28Z
dc.date.issued2014-08en_US
dc.date.submitted2016-06-15T15:10:07.210Z
dc.identifier.urihttp://qmro.qmul.ac.uk/xmlui/handle/123456789/12959
dc.description.abstractBinding and hydrolysis of ATP is universally required by AAA+ proteins to underpin their mechano-chemical work. Here we explore the roles of the ATPase site in an AAA+ transcriptional activator protein, the phage shock protein F (PspF), by specifically altering the Walker B motif sequence required in catalyzing ATP hydrolysis. One such mutant, the E108Q variant, is defective in ATP hydrolysis but fully remodels target transcription complexes, the RNAP-σ(54) holoenzyme, in an ATP dependent manner. Structural analysis of the E108Q variant reveals that unlike wild-type protein, which has distinct conformations for E108 residue in the ATP and ADP bound forms, E108Q adapts the same conformation irrespective of nucleotide bound. Our data show that the remodeling activities of E108Q are strongly favored on pre-melted DNA and engagement with RNAP-σ(54) using ATP binding can be sufficient to convert the inactive holoenzyme to an active form, while hydrolysis per se is required for nucleic acid remodeling that leads to transcription bubble formation. Furthermore, using linked dimer constructs, we show that RNAP-σ(54) engagement by adjacent subunits within a hexamer are required for this protein remodeling activity while DNA remodeling activity can tolerate defective ATP hydrolysis of alternating subunits.en_US
dc.format.extent9249 - 9261en_US
dc.languageengen_US
dc.language.isoenen_US
dc.relation.ispartofNucleic Acids Resen_US
dc.rightsCC-BY
dc.subjectAdenosine Diphosphateen_US
dc.subjectAdenosine Triphosphatasesen_US
dc.subjectAdenosine Triphosphateen_US
dc.subjectDNAen_US
dc.subjectEscherichia coli Proteinsen_US
dc.subjectGlutamic Aciden_US
dc.subjectModels, Molecularen_US
dc.subjectMutationen_US
dc.subjectTrans-Activatorsen_US
dc.subjectTranscription, Geneticen_US
dc.titleMolecular basis of nucleotide-dependent substrate engagement and remodeling by an AAA+ activator.en_US
dc.typeArticle
dc.rights.holder© 2014 The Author(s)
dc.identifier.doi10.1093/nar/gku588en_US
pubs.author-urlhttps://www.ncbi.nlm.nih.gov/pubmed/25063294en_US
pubs.issue14en_US
pubs.notesNot knownen_US
pubs.publication-statusPublisheden_US
pubs.volume42en_US


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