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Figure 1. Intimin-expressing bacteria attach efficiently to mammalian cells expressing membrane-targeted Tir. (A) Full-length Tir (TirFL) and TirMC, a Tir derivative lacking the NH2-terminal cytoplasmic domain, are depicted. TirMC was directed to the plasma membrane by an NH2-terminal targeting sequence derived from a viral HN-protein. Arrows indicate the positions of HA-epitope tags and Y474. (B) HEp-2 cells transfected with plasmids encoding TirFL or TirMC were stained with an mAb to the HA epitope to identify Tir-expressing cells, with an mAb to visualize phosphotyrosine (Y-PO4), and with phalloidin to visualize F-actin. The inset shows several TirFL-expressing cells. (C) Mock-transfected HeLa cells, or cells expressing TirFL, TirMC, or TirMC(Y474F) (identified by anti-HA fluorescence) were challenged with a laboratory strain of E. coli carrying a plasmid-encoding intimin or a pUC19 vector control. Cell binding index, defined as the percentage of cells with at least five bacteria bound, was measured microscopically, and data represent the mean of two experiments. Image published in: Campellone KG et al. (2004) Copyright © 2004, The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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