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J Biol Chem. 2017 Dec 1;292(48):19628-19638. doi: 10.1074/jbc.M117.808394. Epub 2017 Oct 5.

Defining the interaction of the protease CpaA with its type II secretion chaperone CpaB and its contribution to virulence in Acinetobacter species.

Author information

1
From the Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110.
2
the Department of Biological Sciences, University of Alberta, Edmonton T6G 2E9, Alberta, Canada, and.
3
the Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee 37232.
4
From the Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110, mariofeldman@wustl.edu.

Abstract

Acinetobacter baumannii, Acinetobacter nosocomialis, and Acinetobacter pittii are a frequent cause of multidrug-resistant, healthcare-associated infections. Our previous work demonstrated that A. nosocomialis M2 possesses a functional type II secretion system (T2SS) that is required for full virulence. Further, we identified the metallo-endopeptidase CpaA, which has been shown previously to cleave human Factor V and deregulate blood coagulation, as the most abundant type II secreted effector protein. We also demonstrated that its secretion is dependent on CpaB, a membrane-bound chaperone. In this study, we show that CpaA expression and secretion are conserved across several medically relevant Acinetobacter species. Additionally, we demonstrate that deletion of cpaA results in attenuation of A. nosocomialis M2 virulence in moth and mouse models. The virulence defects resulting from the deletion of cpaA were comparable with those observed upon abrogation of T2SS activity. The virulence defects resulting from the deletion of cpaA are comparable with those observed upon abrogation of T2SS activity. We also show that CpaA and CpaB strongly interact, forming a complex in a 1:1 ratio. Interestingly, deletion of the N-terminal transmembrane domain of CpaB results in robust secretion of CpaA and CpaB, indicating that the transmembrane domain is dispensable for CpaA secretion and likely functions to retain CpaB inside the cell. Limited proteolysis of spheroplasts revealed that the C-terminal domain of CpaB is exposed to the periplasm, suggesting that this is the site where CpaA and CpaB interact in vivo Last, we show that CpaB does not abolish the proteolytic activity of CpaA against human Factor V. We conclude that CpaA is, to the best of our knowledge, the first characterized, bona fide virulence factor secreted by Acinetobacter species.

KEYWORDS:

bacterial pathogenesis; bacterial toxin; chaperone; metalloprotease; type II secretion system (T2SS)

PMID:
28982978
PMCID:
PMC5712607
[Available on 2018-12-01]
DOI:
10.1074/jbc.M117.808394
[Indexed for MEDLINE]
Free PMC Article

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