The Gram-negative enteric bacterium Proteus mirabilis is a frequent cause of urinary tract infections (UTIs) in individuals with long-term indwelling catheters or with complicated urinary tracts. The recent release of the P. mirabilis strain HI4320 genome sequence has facilitated identification of potential virulence factors in this organism. Genes appearing to encode a type III secretion system (TTSS) were found in a low GC-content pathogenicity island in the P. mirabilis chromosome. This island contains 24 intact genes that appear to encode all components necessary to assemble a TTSS needle complex, plus at least two putative secreted effector proteins and their chaperones. The genetic organization of the TTSS genes is very similar to that of the TTSS of Shigella flexneri. RT-PCR analysis indicated that these genes are expressed at low levels in vitro. However, insertional mutation of two putative TTSS genes, encoding the requisite ATPase and a possible negative regulator, resulted in no change in either the growth rate of the mutant or the secreted protein profile compared to wild-type. Furthermore, there was no difference in quantitative cultures of urine, bladder and kidney between the ATPase mutant and the wild-type strain in the mouse model of ascending UTI in either independent challenge or co-challenge experiments. The role of the P. mirabilis TTSS, if any, is yet to be determined.