Identification of functional regions within invasion plasmid antigen C (IpaC) of Shigella flexneri.
Shigella flexneri causes bacillary dysentery with symptoms resulting from the inflammation that accompanies bacterial entry into the cells of the colonic epithelium. The effectors of S. flexneri invasion are the Ipa proteins, particularly IpaB and IpaC, which are secreted at the host-pathogen interface following bacterial contact with a host cell. Of the purified Ipa proteins, only IpaC has been shown to possess quantifiable in vitro activities that are related to cellular invasion. In this study, ipaC deletion mutants were generated to identify functional regions within the IpaC protein. From these data, we now know that the N-terminus and an immunogenic central region are not required for IpaC-dependent enhancement of cellular invasion by S. flexneri. However, to restore invasiveness to an ipaC null mutant of S. flexneri, the N-terminus is essential, because IpaC mutants lacking the N-terminus are not secreted by the bacterium. Deletion of the central hydrophobic region eliminates IpaC's ability to interact with phospholipid membranes, and fusion of this region to a modified form of green fluorescent protein converts it into an efficient membrane-associating protein. Meanwhile, deletion of the C-terminus eliminates the mutant protein's ability to establish protein-protein contacts with full-length IpaC. Interestingly, the mutant form of ipaC that restores partial invasiveness to the S. flexneri ipaC null mutant also restores full contact-mediated haemolysis activity to this bacterium. These data support a model in which IpaC possesses a distinct functional organization that is important for bacterial invasion. This information will be important in defining the precise role of IpaC in S. flexneri pathogenesis and in exploring the potential effects of purified IpaC at mucosal surfaces.
Picking
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WL
Department of Molecular Biosciences, University of Kansas, 8047 Haworth, Lawrence, KS 66045, USA. picking@eagle.cc.ukans.edu
Coye
L
L
Osiecki
J C
JC
Barnoski Serfis
A
A
Schaper
E
E
Picking
W D
WD
D000942:Antigens, Bacterial / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000382:microbiology; D005798:Genes, Bacterial; D006461:Hemolysis / Q000235:genetics; D009154:Mutation; D010743:Phospholipids / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D011485:Protein Binding; D017384:Sequence Deletion; D012763:Shigella flexneri / Q000235:genetics / Q000472:pathogenicity*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D010743:Phospholipids; C092342:IpaC protein, Shigella