Characterization of the putative type III secretion ATPase CdsN (Cpn0707) of Chlamydophila pneumoniae.

Journal of Bacteriology
Chris B StoneJames B Mahony

Abstract

Type III secretion (T3S) is utilized by a wide range of gram-negative bacterial pathogens to allow the efficient delivery of effector proteins into the host cell cytoplasm through the use of a syringe-like injectisome. Chlamydophila pneumoniae is a gram-negative, obligate intracellular pathogen that has the structural genes coding for a T3S system, but the functionality of the system has not yet been demonstrated. T3S is dependent on ATPase activity, which catalyzes the unfolding of proteins and the secretion of effector proteins through the injectisome. CdsN (Cpn0707) is predicted to be the T3S ATPase of C. pneumoniae based on sequence similarity to other T3S ATPases. Full-length CdsN and a C-terminal truncation of CdsN were cloned as glutathione S-transferase (GST)-tagged constructs and expressed in Escherichia coli. The GST-tagged C-terminal truncation of CdsN possessed ATPase activity, catalyzing the release of ADP and P(i) from ATP at a rate of 0.55 +/- 0.07 micromol min(-1) mg(-1) in a time- and dose-dependent manner. CdsN formed oligomers and high-molecular-weight multimers, as assessed by formaldehyde fixation and nondenaturing polyacrylamide gel electrophoresis. Using bacterial two-hybrid and GST pull-down assays, CdsN...Continue Reading

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Citations

Jan 26, 2010·BMC Microbiology·Chris B StoneJames B Mahony
Oct 31, 2015·Microbes and Infection·Joshua C Ferrell, Kenneth A Fields
Jan 18, 2015·International Journal of Medical Microbiology : IJMM·Julia KästnerFrank Hänel
Mar 8, 2016·Protein Science : a Publication of the Protein Society·Jamie L BurgessNicholas E Dickenson
Jan 13, 2015·Experimental and Therapeutic Medicine·Yuyu ChenQiugui Zhang
Feb 18, 2009·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·J M KernM Maass
Jul 19, 2014·The Journal of Biological Chemistry·Sarah E AllisonBrian K Coombes
May 6, 2011·The Journal of Immunology : Official Journal of the American Association of Immunologists·Norma Olivares-ZavaletaHarlan D Caldwell

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