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Intracellular survival of Yersinia spp. in macrophages requires outer membrane protein A (OmpA)
Umeå University, Faculty of Science and Technology, Department of Molecular Biology (Faculty of Science and Technology). (Hans Wolf-Watz)
Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA.
Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA.
Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine, Miami, Florida, USA.
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(English)Manuscript (preprint) (Other academic)
Abstract [en]

In order to grow, multiply and disseminate within their hosts, Yersinia spp. must be able tosurvive encounters with host phagocytic cells such as macrophages. Yersinia pestis and Yersinia pseudotuberculosis both have the ability to survive within the phagolysosome of macrophages which may be important in early stages of the infection. These organisms also employ a T3SS toactively prevent phagocytosis. The ability of Y. pestis KIM mutants defective in the expression of outer membrane proteins to survive and proliferate in the presence of RAW 264.7 macrophage-like cells was evaluated. A Y. pestis KIM ΔompA mutant exhibited a significant defect in proliferation in the presence of macrophages. A similar defect could be produced by overexpressing the negative regulatory sRNA MicA, which down regulates OmpA expression. The ΔompA mutant exhibited no defect in the T3SS-mediated secretion or injection of Yops; however, this mutant showed a defect in intra macrophage survival in gentamicin protection assays. These studies suggest that the Yersinia spp. OmpA protein is a virulence factor that is required for survival within macrophages.

National Category
Microbiology in the medical area
Identifiers
URN: urn:nbn:se:umu:diva-33952OAI: oai:DiVA.org:umu-33952DiVA: diva2:318876
Available from: 2010-05-11 Created: 2010-05-11 Last updated: 2010-06-03Bibliographically approved
In thesis
1. Outer membrane proteins of Yersinia pestis: Ail and OmpA
Open this publication in new window or tab >>Outer membrane proteins of Yersinia pestis: Ail and OmpA
2010 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

A vast number of studies have been completed on the virulence determinants of Yersinia spp.; however, the focus of many of these studies has been on the virulence plasmid and the plasmid-encoded Type three secretion system. Nevertheless, many chromosomal genes whose products are directly involved in virulence have also been identified. Some of these critical virulence determinants are outer membrane proteins. Outer membrane proteins of Gram-negative bacteria often have important physiological roles; however, some have also been found to be important for pathogenesis. In this thesis, we investigated two Yersinia. pestis outer membrane proteins, Ail and OmpA, and their roles in virulence. We provide evidence that Y. pestis Ail is a highly expressed outer membrane protein that is absolutely essential for Y. pestis to resist the killing action of the complement system present in human blood and tissues, as well as the blood and tissues of other mammalian hosts. Furthermore, Ail was important for virulence in a Y. pestis-Canorhabditis elegans model of infection.The work in this thesis also provided the first evidence that another surface-exposed outer membrane protein, termed OmpA, is required for both Yersinia pseudotuberculosis and Y. pestis to survive and proliferate intracellularly in macrophages. Finally, we provide evidence that Y. pestis has a functional small RNA MicA that controls the expression of OmpA. This is the first demonstration of sRNA-mediated regulation of a Yersinia virulence factor. This work has paved the way for future studies on the role of outer membrane proteins in virulence, particularly the role of Ail and OmpA.

Place, publisher, year, edition, pages
Umeå: Institutionen för molekylärbiologi (Teknisk-naturvetenskaplig fakultet), Umeå universitet, 2010. 89 p.
Series
Doctoral thesis / Umeå University, Department of Molecular Biology
Keyword
Yersinia pestis, outer membrane proteins, Ail, ompA
National Category
Microbiology in the medical area
Research subject
Immunology; Microbiology; Infectious Diseases
Identifiers
urn:nbn:se:umu:diva-33956 (URN)978-91-7459-030-2 (ISBN)
Public defence
2010-06-03, Molekylärbiologi, Major Groove, Byggnad 6L, Umeå universitet, Umeå, 10:00 (English)
Opponent
Supervisors
Available from: 2010-05-12 Created: 2010-05-11 Last updated: 2010-05-18Bibliographically approved

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