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Article: Covalently linking the Escherichia coli global anaerobic regulator FNR in tandem allows it to function as an oxygen stable dimer

TitleCovalently linking the Escherichia coli global anaerobic regulator FNR in tandem allows it to function as an oxygen stable dimer
Authors
KeywordsEMSA
FNR
Transcription regulator
Issue Date2012
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/description
Citation
Biochemical And Biophysical Research Communications, 2012, v. 419 n. 1, p. 43-48 How to Cite?
AbstractThe anaerobic global regulator FNR from Escherichia coli is a [4Fe-4S] 2+ cluster containing, O 2 labile dimer that plays an important role in adapting the bacterium to its anaerobic lifestyle. Although functional significance of this global regulator has been well established, its structural and biochemical characterizations have been hindered by the intrinsic O 2 lability of FNR protein. To obtain oxygen stable FNR variants for in vitro characterizations, in this study, we utilized the approach of covalently linking two fnr alleles in tandem to promote their in situ dimerization in the absence of the [4Fe-4S] 2+ cluster under aerobic conditions. By covalently linking two alleles of fnrD154A mutant, which has reduced charge repulsions between two FNR monomers, an FNR variant of FNRD154A-Linker-FNRD154A (designated as (FNRD154A) 2) was constructed. (FNRD154A) 2 displayed significantly enhanced DNA binding affinities and transcriptional regulatory activities to various FNR dependent promoters under aerobic conditions. Further in vivo and in vitro studies demonstrated that this variant retains oxygen sensing capability and maintains a similar secondary structure as that formed by native monomers of FNRD154A. We conclude that this novel variant of FNR can be widely used in various biochemical and structural studies of FNR in the presence of O 2. © 2012 Elsevier Inc.
Persistent Identifierhttp://hdl.handle.net/10722/160583
ISSN
2015 Impact Factor: 2.371
2015 SCImago Journal Rankings: 1.152
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorShan, Yen_HK
dc.contributor.authorPan, Qen_HK
dc.contributor.authorLiu, Jen_HK
dc.contributor.authorHuang, Fen_HK
dc.contributor.authorSun, Hen_HK
dc.contributor.authorNishino, Ken_HK
dc.contributor.authorYan, Aen_HK
dc.date.accessioned2012-08-16T06:14:46Z-
dc.date.available2012-08-16T06:14:46Z-
dc.date.issued2012en_HK
dc.identifier.citationBiochemical And Biophysical Research Communications, 2012, v. 419 n. 1, p. 43-48en_HK
dc.identifier.issn0006-291Xen_HK
dc.identifier.urihttp://hdl.handle.net/10722/160583-
dc.description.abstractThe anaerobic global regulator FNR from Escherichia coli is a [4Fe-4S] 2+ cluster containing, O 2 labile dimer that plays an important role in adapting the bacterium to its anaerobic lifestyle. Although functional significance of this global regulator has been well established, its structural and biochemical characterizations have been hindered by the intrinsic O 2 lability of FNR protein. To obtain oxygen stable FNR variants for in vitro characterizations, in this study, we utilized the approach of covalently linking two fnr alleles in tandem to promote their in situ dimerization in the absence of the [4Fe-4S] 2+ cluster under aerobic conditions. By covalently linking two alleles of fnrD154A mutant, which has reduced charge repulsions between two FNR monomers, an FNR variant of FNRD154A-Linker-FNRD154A (designated as (FNRD154A) 2) was constructed. (FNRD154A) 2 displayed significantly enhanced DNA binding affinities and transcriptional regulatory activities to various FNR dependent promoters under aerobic conditions. Further in vivo and in vitro studies demonstrated that this variant retains oxygen sensing capability and maintains a similar secondary structure as that formed by native monomers of FNRD154A. We conclude that this novel variant of FNR can be widely used in various biochemical and structural studies of FNR in the presence of O 2. © 2012 Elsevier Inc.en_HK
dc.languageengen_US
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/descriptionen_HK
dc.relation.ispartofBiochemical and Biophysical Research Communicationsen_HK
dc.subjectEMSAen_HK
dc.subjectFNRen_HK
dc.subjectTranscription regulatoren_HK
dc.subject.meshCodon, Terminator - genetics-
dc.subject.meshEscherichia coli - genetics - metabolism-
dc.subject.meshEscherichia coli Proteins - chemistry - genetics - metabolism-
dc.subject.meshIron-Sulfur Proteins - chemistry - genetics - metabolism-
dc.subject.meshOxygen - chemistry - metabolism-
dc.titleCovalently linking the Escherichia coli global anaerobic regulator FNR in tandem allows it to function as an oxygen stable dimeren_HK
dc.typeArticleen_HK
dc.identifier.emailSun, H: hsun@hku.hken_HK
dc.identifier.emailYan, A: ayan8@hku.hken_HK
dc.identifier.authoritySun, H=rp00777en_HK
dc.identifier.authorityYan, A=rp00823en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.bbrc.2012.01.121en_HK
dc.identifier.pmid22326866-
dc.identifier.scopuseid_2-s2.0-84862821325en_HK
dc.identifier.hkuros203635en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-84862821325&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume419en_HK
dc.identifier.issue1en_HK
dc.identifier.spage43en_HK
dc.identifier.epage48en_HK
dc.identifier.isiWOS:000301560500008-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridShan, Y=55262302600en_HK
dc.identifier.scopusauthoridPan, Q=55262239500en_HK
dc.identifier.scopusauthoridLiu, J=55261291900en_HK
dc.identifier.scopusauthoridHuang, F=55261648500en_HK
dc.identifier.scopusauthoridSun, H=7404827446en_HK
dc.identifier.scopusauthoridNishino, K=7102938399en_HK
dc.identifier.scopusauthoridYan, A=8621667000en_HK
dc.identifier.citeulike10326075-

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