ApoFnr binds as a monomer to promoters regulating expression of enterotoxin genes of Bacillus cereus. - Archive ouverte HAL
Article Dans Une Revue Journal of Bacteriology Année : 2008

ApoFnr binds as a monomer to promoters regulating expression of enterotoxin genes of Bacillus cereus.

Résumé

Bacillus cereus Fnr is a member of the Crp/Fnr (cAMP-binding protein/fumarate nitrate reduction regulatory protein) family of helix-turn-helix transcriptional regulators. It is essential for the expression of Hbl and Nhe enterotoxin genes independently of the oxygen tension in the environment. We studied aerobic Fnr binding to target sites in promoters regulating the expression of enterotoxin genes. B. cereus Fnr was overexpressed and purified as either a C-terminal His-tagged (FnrHis) fusion protein or an N-terminal fusion protein tagged with the Strep-tag (IBA BioTAGnology) (StrepFnr). Both recombinant Fnr proteins were produced as apoforms (clusterless) and occured as mixtures of monomers and oligomers in solution. However, apoFnrHis was mainly monomeric, while apoStrepFnr was mainly oligomeric, suggesting that the His-tagged C-terminal extremity may interfere with oligomerization. The oligomeric state of apoStrepFnr was dithiothreitol sensitive, underlining the importance of a disulphide bridge for apoFnr oligomerization. Electrophoretic mobility shift assays showed that monomeric apoFnr, but not oligomeric apoFnr, bound to specific sequences located in the promoter regions of the enterotoxin regulators fnr, resDE and plcR and the structural genes hbl and nhe. The question of whether apoFnr binding is regulated in vivo by redox-dependent oligomerization is discussed.
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Dates et versions

hal-00378324 , version 1 (24-04-2009)

Identifiants

  • HAL Id : hal-00378324 , version 1
  • PRODINRA : 249771

Citer

Julia Esbelin, Yves Jouanneau, J. Armengaud, Catherine Duport. ApoFnr binds as a monomer to promoters regulating expression of enterotoxin genes of Bacillus cereus.. Journal of Bacteriology, 2008, 190, pp.4242-4251. ⟨hal-00378324⟩
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