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Detailed analysis of Helicobacter pylori Fur‐regulated promoters reveals a Fur box core sequence and novel Fur‐regulated genes
Author(s) -
Pich Oscar Q.,
Carpenter Beth M.,
Gilbreath Jeremy J.,
Merrell D. Scott
Publication year - 2012
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.2012.08066.x
Subject(s) - promoter , biology , gene , genetics , repressor , microbiology and biotechnology , gene expression
Summary In Helicobacter pylori , iron balance is controlled by the Ferric uptake regulator (Fur), an iron‐sensing repressor protein that typically regulates expression of genes implicated in iron transport and storage. Herein, we carried out extensive analysis of Fur‐regulated promoters and identified a 7‐1‐7 motif with dyad symmetry (5′‐TAATAATnATTATTA‐3′), which functions as the Fur box core sequence of H. pylori . Addition of this sequence to the promoter region of a typically non‐Fur regulated gene was sufficient to impose Fur‐dependent regulation in vivo . Moreover, mutation of this sequence within Fur‐controlled promoters negated regulation. Analysis of the H. pylori chromosome for the occurrence of the Fur box established the existence of well‐conserved Fur boxes in the promoters of numerous known Fur‐regulated genes, and revealed novel putative Fur targets. Transcriptional analysis of the new candidate genes demonstrated Fur‐dependent repression of HPG27_51, HPG27_52, HPG27_199, HPG27_445, HPG27_825 and HPG27_1063, as well as Fur‐mediated activation of the cytotoxin associated gene A, cagA (HPG27_507). Furthermore, electrophoretic mobility shift assays confirmed specific binding of Fur to the promoters of each of these genes. Future experiments will determine whether loss of Fur regulation of any of these particular genes contributes to the defects in colonization exhibited by the H. pylori fur mutant.

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