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To identify adhesins critical for Yop delivery, we initiated two transposon mutagenesis screens using the<jats:italic>mariner<\/jats:italic>transposon. To avoid redundant cell binding activities, we initiated the screen with a strain deleted for two known adhesins, pH 6 antigen and the autotransporter, YapC, as well as the Caf1 capsule, which is known to obscure some adhesins. The mutants that emerged contained insertions within the<jats:italic>ail<\/jats:italic>(attachment and invasion locus) gene of<jats:italic>Y. pestis<\/jats:italic>. A reconstructed mutant with a single deletion in the<jats:italic>ail<\/jats:italic>locus (y1324) was severely defective for delivery of Yops to HEp-2 human epithelial cells and significantly defective for delivery of Yops to THP-1 human monocytes. Specifically, the Yop delivery defect was apparent when cell rounding and translocation of an ELK-tagged YopE derivative into host cells were monitored. Although the<jats:italic>ail<\/jats:italic>mutant showed only a modest decrease in cell binding capacity in vitro, the KIM5 \u0394<jats:italic>ail<\/jats:italic>mutant exhibited a &gt;3,000-fold-increased 50% lethal dose in mice. Mice infected with the \u0394<jats:italic>ail<\/jats:italic>mutant also had 1,000-fold fewer bacteria in their spleens, livers, and lungs 3 days after infection than did those infected with the parental strain, KIM5. Thus, the Ail protein is critical for both<jats:italic>Y. pestis<\/jats:italic>type III secretion in vitro and infection in mice.<\/jats:p>","DOI":"10.1128\/iai.00913-08","type":"journal-article","created":{"date-parts":[[2008,12,9]],"date-time":"2008-12-09T01:25:40Z","timestamp":1228785940000},"page":"825-836","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":88,"title":["The<i>Yersinia pestis<\/i>Ail Protein Mediates Binding and Yop Delivery to Host Cells Required for Plague Virulence"],"prefix":"10.1128","volume":"77","author":[{"given":"Suleyman","family":"Felek","sequence":"first","affiliation":[{"name":"Department of Biologic and Materials Sciences, University of Michigan School of Dentistry"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eric S.","family":"Krukonis","sequence":"additional","affiliation":[{"name":"Department of Biologic and Materials Sciences, University of Michigan School of Dentistry"},{"name":"Department of Microbiology and Immunology, University of Michigan School of Medicine, Ann Arbor, Michigan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.23.13658"},{"key":"e_1_3_2_3_2","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1046\/j.1365-2958.2001.02676.x","volume":"42","year":"2001","unstructured":"Alrutz, M. 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