%0 Journal Article %A Bridge, Dacie R. %A Novotny, Matthew J. %A Moore, Elizabeth R. %A Olson, Joan C. %T Role of host cell polarity and leading edge properties in Pseudomonas type III secretion %D 2010 %J Microbiology, %V 156 %N 2 %P 356-373 %@ 1465-2080 %R https://doi.org/10.1099/mic.0.033241-0 %K CTB, cholera toxin B subunit %K ROCK, Rho kinase %K HA, haemagglutinin %K Pa-T3S, P. aeruginosa type III secretion %K MT, microtubule %K dsPFO, prepore locked PFO %K MβCD, methyl-beta-cyclodextrin %K IQGAP1, IQ-motif-containing GTPase-activating protein 1 %K TPA, 12-O-tetradeconylphorbol-13-acetate %K PFO, perfringolysin O %K LatB, latrunculin B %K Pa-ExoS-HA, P. aeruginosa strain PA103ΔUT expressing plasmid-encoded ExoS-HA %K siRNA, small interfering RNA %K IF, immunofluorescent/immunofluorescence %K ssPFO, monomer locked PFO %K T3S, type III secretion %K LE, leading edge %I Microbiology Society, %X Type III secretion (T3S) functions in establishing infections in a large number of Gram-negative bacteria, yet little is known about how host cell properties might function in this process. We used the opportunistic pathogen Pseudomonas aeruginosa and the ability to alter host cell sensitivity to Pseudomonas T3S to explore this problem. HT-29 epithelial cells were used to study cellular changes associated with loss of T3S sensitivity, which could be induced by treatment with methyl-beta-cyclodextrin or perfringolysin O. HL-60 promyelocytic cells are innately resistant to Pseudomonas T3S and were used to study cellular changes occurring in response to induction of T3S sensitivity, which occurred following treatment with phorbol esters. Using both cell models, a positive correlation was observed between eukaryotic cell adherence to tissue culture wells and T3S sensitivity. In examining the type of adhesion process linked to T3S sensitivity in HT-29 cells, a hierarchical order of protein involvement was identified that paralleled the architecture of leading edge (LE) focal complexes. Conversely, in HL-60 cells, induction of T3S sensitivity coincided with the onset of LE properties and the development of actin-rich projections associated with polarized cell migration. When LE architecture was examined by immunofluorescent staining for actin, Rac1, IQ-motif-containing GTPase-activating protein 1 (IQGAP1) and phosphatidylinositol 3 kinase (PI3 kinase), intact LE structure was found to closely correlate with host cell sensitivity to P. aeruginosa T3S. Our model for host cell involvement in Pseudomonas T3S proposes that cortical actin polymerization at the LE alters membrane properties to favour T3S translocon function and the establishment of infections, which is consistent with Pseudomonas infections targeting wounded epithelial barriers undergoing cell migration. %U https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.033241-0