Specificity of NifEN and VnfEN for the Assembly of Nitrogenase Active Site Cofactors in Azotobacter vinelandii

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dc.contributor.authorPerez-Gonzalez, Anaen
dc.contributor.authorJimenez-Vicente, Emilioen
dc.contributor.authorGies-Elterlein, Jakoben
dc.contributor.authorSalinero-Lanzarote, Alvaroen
dc.contributor.authorYang, Zhi-Yongen
dc.contributor.authorEinsle, Oliveren
dc.contributor.authorSeefeldt, Lance C.en
dc.contributor.authorDean, Dennis R.en
dc.date.accessioned2022-01-17T14:45:41Zen
dc.date.available2022-01-17T14:45:41Zen
dc.date.issued2021-07-01en
dc.date.updated2022-01-17T14:45:36Zen
dc.description.abstractThe nitrogen-fixing microbe Azotobacter vinelandii has the ability to produce three genetically distinct, but mechanistically similar, components that catalyze nitrogen fixation. For two of these components, the Mo-dependent and V-dependent components, their corresponding metal-containing active site cofactors, designated FeMo-cofactor and FeV-cofactor, respectively, are preformed on separate molecular scaffolds designated NifEN and VnfEN, respectively. From prior studies, and the present work, it is now established that neither of these scaffolds can replace the other with respect to their in vivo cofactor assembly functions. Namely, a strain inactivated for NifEN cannot produce active Mo-dependent nitrogenase nor can a strain inactivated for VnfEN produce an active V-dependent nitrogenase. It is therefore proposed that metal specificities for FeMo-cofactor and FeV-cofactor formation are supplied by their respective assembly scaffolds. In the case of the third, Fe-only component, its associated active site cofactor, designated FeFe-cofactor, requires neither the NifEN nor VnfEN assembly scaffold for its formation. Furthermore, there are no other genes present in A. vinelandii that encode proteins having primary structure similarity to either NifEN or VnfEN. It is therefore concluded that FeFe-cofactor assembly is completed within its cognate catalytic protein partner without the aid of an intermediate assembly site. IMPORTANCE Biological nitrogen fixation is a complex process involving the nitrogenases. The biosynthesis of an active nitrogenase involves a large number of genes and the coordinated function of their products. Understanding the details of the assembly and activation of the different nitrogen fixation components, in particular the simplest one known so far, the Fe-only nitrogenase, would contribute to the goal of transferring the necessary genetic elements of bacterial nitrogen fixation to cereal crops to endow them with the capacity for self-fertilization. In this work, we show that there is no need for a scaffold complex for the assembly of the FeFecofactor, which provides the active site for Fe-only nitrogenase. These results are in agreement with previously reported genetic reconstruction experiments using a non-nitrogen-fixing microbe. In aggregate, these findings provide a high degree of confidence that the Fe-only system represents the simplest and, therefore, most attractive target for mobilizing nitrogen fixation into plants.en
dc.description.versionPublished versionen
dc.format.extent16 page(s)en
dc.format.mimetypeapplication/pdfen
dc.identifierARTN e01568-21 (Article number)en
dc.identifier.doihttps://doi.org/10.1128/mBio.01568-21en
dc.identifier.eissn2150-7511en
dc.identifier.issn2150-7511en
dc.identifier.issue4en
dc.identifier.orcidDean, Dennis [0000-0001-8960-6196]en
dc.identifier.pmid34281397en
dc.identifier.urihttp://hdl.handle.net/10919/107692en
dc.identifier.volume12en
dc.language.isoenen
dc.publisherAmerican Society for Microbiologyen
dc.relation.urihttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000696627000010&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1en
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectLife Sciences & Biomedicineen
dc.subjectMicrobiologyen
dc.subjectassemblyen
dc.subjectFeFe-cofactoren
dc.subjectFeMo-cofactoren
dc.subjectFeV-cofactoren
dc.subjectmolybdenumen
dc.subjectnitrogenaseen
dc.subjectvanadiumen
dc.subjectIRON-MOLYBDENUM COFACTORen
dc.subjectIN-VITRO SYNTHESISen
dc.subjectVANADIUM NITROGENASEen
dc.subjectSTRUCTURAL GENESen
dc.subjectMUTATIONAL ANALYSISen
dc.subjectTRANSCRIPTIONAL REGULATIONen
dc.subjectNUCLEOTIDE-SEQUENCEen
dc.subjectONLY NITROGENASEen
dc.subjectN-2 REDUCTIONen
dc.subjectFEMO-COFACTORen
dc.subject0605 Microbiologyen
dc.subject.meshAzotobacter vinelandiien
dc.subject.meshNitrogenen
dc.subject.meshCoenzymesen
dc.subject.meshNitrogenaseen
dc.subject.meshMolybdoferredoxinen
dc.subject.meshBacterial Proteinsen
dc.subject.meshNitrogen Fixationen
dc.subject.meshCatalytic Domainen
dc.subject.meshBiocatalysisen
dc.titleSpecificity of NifEN and VnfEN for the Assembly of Nitrogenase Active Site Cofactors in Azotobacter vinelandiien
dc.title.serialmBioen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
dc.type.otherArticleen
dc.type.otherJournalen
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciencesen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/Biochemistryen
pubs.organisational-group/Virginia Tech/University Distinguished Professorsen
pubs.organisational-group/Virginia Tech/University Research Institutesen
pubs.organisational-group/Virginia Tech/University Research Institutes/Fralin Life Sciencesen
pubs.organisational-group/Virginia Tech/Faculty of Health Sciencesen
pubs.organisational-group/Virginia Tech/All T&R Facultyen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/CALS T&R Facultyen
pubs.organisational-group/Virginia Tech/University Research Institutes/Fralin Life Sciences/Durelle Scotten

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