Open AccessHP0958 is an essential motility gene in Helicobacter pylori
Author(s) -
Ryan Kieran A.,
Karim Najma,
Worku Mulugeta,
Moore Stanley A.,
Penn Charles W.,
O'Toole Paul W.
Publication year - 2005
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/j.femsle.2005.05.022
Subject(s) - flagellin , rpon , biology , flagellum , mutant , gene , genetics , transcription (linguistics) , microbiology and biotechnology , gene expression , promoter , linguistics , philosophy
Motility is an essential colonization factor for the human gastric pathogen Helicobacter pylori . The H. pylori genome encodes most known flagellar proteins, although a number of key transcription regulators, chaperones, and structural proteins have not yet been identified. Using recently published yeast two‐hybrid data we identified HP0958 as a potential motility‐associated protein due to its strong interactions with RpoN (σ 54 ) and FliH, a flagellar ATPase regulator. HP0958 exhibits no sequence similarity to any published flagellar genes but contains a carboxy‐terminal zinc finger domain that could function in nucleic acid or protein binding. We created a HP0958 mutant by inserting a chloramphenicol resistance marker into the gene using a PCR‐based allelic exchange method and the resultant mutant was non‐motile as measured by a BacTracker instrument. Electron microscopic analysis revealed that the HP0958 mutant cells were aflagellate and Western blot analysis revealed a dramatic reduction in flagellin and hook protein production. The HP0958 mutant also showed decreased transcription of flgE , flaB and flaA as well as the checkpoint genes flhA and flhF . Expression of flgM was increased relative to the wild‐type and both rpoN and fliA (σ 28 ) expression were unchanged. We conclude that HP0958 is essential for normal motility and flagella production, and represents a novel flagellar component in the epsilon proteobacteria.