Reactions of nitric oxide and oxygen with the regulator of fumarate and nitrate reduction, a global transcriptional regulator, during anaerobic growth of Escherichia coli

Methods in Enzymology
Jason C CrackAdrian J Jervis

Abstract

The Escherichia coli fumarate and nitrate reductase (FNR) regulator protein is an important transcriptional regulator that controls the expression of a large regulon of more than 100 genes in response to changes in oxygen availability. FNR is active when it acquires a [4Fe-4S](2+) cluster under anaerobic conditions. The presence of the [4Fe-4S](2+) cluster promotes protein dimerization and site-specific DNA binding, facilitating activation or repression of target promoters. Oxygen is sensed by the controlled disassembly of the [4Fe-4S](2+) cluster, ultimately resulting in inactive, monomeric, apo-FNR. The FNR [4Fe-4S](2+) cluster is also sensitive to nitric oxide, such that under anaerobic conditions the protein is inactivated by nitrosylation of the iron-sulfur cluster, yielding a mixture of monomeric and dimeric dinitrosyl-iron cysteine species. This chapter describes some of the methods used to produce active [4Fe-4S] FNR protein and investigates the reaction of the [4Fe-4S](2+) cluster with nitric oxide and oxygen in vitro.

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Citations

Mar 6, 2009·Proceedings of the National Academy of Sciences of the United States of America·Adrian J JervisJeffrey Green
Mar 9, 2013·The Journal of Biological Chemistry·Jason C CrackNick E Le Brun
Sep 25, 2014·Radiation and Environmental Biophysics·Filipe FolgosaAlice S Pereira
Dec 24, 2017·Nature Communications·Bassam K KudhairJeffrey Green
Nov 21, 2015·Chemistry : a European Journal·Marta K BruskaMarkus Reiher
Aug 12, 2014·Applied and Environmental Microbiology·Anna MühligStefanie Müller-Herbst
Nov 10, 2012·Dalton Transactions : an International Journal of Inorganic Chemistry·Gottfried UndenMareike Singenstreu
May 15, 2015·The Journal of Biological Chemistry·Susan A IbrahimJeffrey Green
Apr 19, 2020·The Journal of Biological Chemistry·Melissa Y Y StewartNick E Le Brun

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