61644 20200221144320.0 doi 10.1089/ars.2014.6175 sideral 99592 ART-2016-99592 eng (orcid)0000-0002-9312-1519 Botello-Morte,L Cysteine mutational studies provide insight into a thiol-based redox switch mechanism of metal and DNA binding in FurA from Anabaena sp. PCC 7120 2016 Access copy available to the general public Unrestricted Aims: The ferric uptake regulator (Fur) is the main transcriptional regulator of genes involved in iron homeostasis in most prokaryotes. FurA from Anabaena sp. PCC 7120 contains five cysteine residues, four of them arranged in two redox-active CXXC motifs. The protein needs not only metal but also reducing conditions to remain fully active in vitro. Through a mutational study of the cysteine residues present in FurA, we have investigated their involvement in metal and DNA binding. Results: Residue C101 that belongs to a conserved CXXC motif plays an essential role in both metal and DNA binding activities in vitro. Substitution of C101 by serine impairs DNA and metal binding abilities of FurA. Isothermal titration calorimetry measurements show that the redox state of C101 is responsible for the protein ability to coordinate the metal corepressor. Moreover, the redox state of C101 varies with the presence or absence of C104 or C133, suggesting that the environments of these cysteines are mutually interdependent. Innovation: We propose that C101 is part of a thiol/disulfide redox switch that determines FurA ability to bind the metal corepressor. Conclusion: This mechanism supports a novel feature of a Fur protein that emerges as a regulator, which connects the response to changes in the intracellular redox state and iron management in cyanobacteria. info:eu-repo/grantAgreement/ES/DGA/B89 info:eu-repo/grantAgreement/ES/FIS/PI10-00186 info:eu-repo/grantAgreement/ES/ISCIII/CP07-00289 info:eu-repo/grantAgreement/ES/MICINN/BFU2010-19451 info:eu-repo/grantAgreement/ES/MINECO/BFU2012-31458 info:eu-repo/grantAgreement/ES/MINECO/CSD2008-00005 info:eu-repo/grantAgreement/ES/MINECO/CTQ2011-24393 info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 6.337 2016 ENDOCRINOLOGY & METABOLISM 13 / 138 = 0.094 2016 Q1 T1 BIOCHEMISTRY & MOLECULAR BIOLOGY 34 / 287 = 0.118 2016 Q1 T1 2.896 2016 Biochemistry 2016 Q1 Cell Biology 2016 Q1 Physiology 2016 Q1 Medicine (miscellaneous) 2016 Q1 Molecular Biology 2016 Q1 Clinical Biochemistry 2016 Q1 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Pellicer,S. Contreras,L. M. Neira,J. L. (orcid)0000-0001-5664-1729 Abian,O. Universidad de Zaragoza (orcid)0000-0001-5702-4538 Velazquez-Campoy,A. Universidad de Zaragoza (orcid)0000-0002-2742-3711 Peleato,M. L. Universidad de Zaragoza (orcid)0000-0001-8644-4574 Fillat,M. F. Universidad de Zaragoza (orcid)0000-0002-8181-2689 Bes,M. T. Universidad de Zaragoza 1002 412 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Fisiología Vegetal 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 24, 4 (2016), 173-185 Antioxid. redox signal. ANTIOXIDANTS & REDOX SIGNALING 1523-0864 880572 http://zaguan.unizar.es/record/61644/files/texto_completo.pdf Versión publicada 116219 http://zaguan.unizar.es/record/61644/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:61644 articulos driver 2020-02-21-13:41:16 ARTICLE