IcmR-regulated membrane insertion and efflux by the Legionella pneumophila IcmQ protein.
Legionella pneumophila proliferates within alveolar macrophages as a central property of Legionnaires' disease. Intracellular growth involves formation of a replicative phagosome, which requires the bacterial Dot/Icm system, a multiprotein secretion apparatus that translocates proteins from the bacterium across the macrophage plasma membrane. Two components of this system, IcmR and IcmQ, are proposed to exhibit a chaperone/substrate relationship similar to that observed in other protein translocation systems. We report here that IcmQ inserts into lipid membranes and forms pores that allow the efflux of the dye calcein but not Dextran 3000. Both membrane insertion and pore formation were inhibited by IcmR. Trypsin digestion mapping demonstrated that IcmQ is subdivided into two functional domains. The N-terminal region of IcmQ was necessary and sufficient for insertion into lipid membranes and calcein efflux. The C-terminal domain was necessary for efficient association of the protein with lipid bilayers. IcmR was found to bind to the N-terminal portion of the protein thus providing a mechanism for its ability to inhibit IcmQ pore-forming activity. Localization of IcmQ on the surface of the L. pneumophila shortly after infection as well as its pore-forming capacities suggest a role for IcmQ in forming a channel that leads translocated effectors out of the bacterium.
Duménil
Guillaume
G
Howard Hughes Medical Institute, Boston, Massachusetts 02111, USA.
Montminy
Timothy P
TP
Tang
May
M
Isberg
Ralph R
RR
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001693:Biological Transport, Active; D002462:Cell Membrane / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D016952:Legionella pneumophila / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D008264:Macrophages / Q000382:microbiology; D008563:Membrane Lipids / Q000378:metabolism; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C432955:ICMQ protein, Legionella pneumophila; C432954:IcmR protein, Legionella pneumophila; D008563:Membrane Lipids; D018832:Molecular Chaperones