期刊
EUROPEAN JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
卷 5, 期 1, 页码 68-80出版社
AKADEMIAI KIADO ZRT
DOI: 10.1556/EuJMI-D-15-00003
关键词
HtrA; secretion; chaperone; flagellum; E-cadherin; molecular pathogenesis; cellular invasion; signaling; transwell; paracellular; virulence; stress response
类别
Campylobacter jejuni is an important pathogen of foodborne illness. Transmigration across the intestinal epithelial barrier and invasion are considered as primary reasons for tissue damage triggered by C. jejuni. Using knockout mutants, it was shown that the serine protease HtrA may be important for stress tolerance and physiology of C. jejuni. HtrA is also secreted in the extracellular environment, where it can cleave junctional host cell proteins such as E-cadherin. Aim of the present study was to establish a genetic complementation system in two C. jejuni strains in order to introduce the wild-type htrA gene in trans, test known htrA phenotypes, and provide the basis to perform further mutagenesis. We confirm that reexpression of the htrA wild-type gene in Delta htrA mutants restored the following phenotypes: 1) C. jejuni growth at high temperature (44 degrees C), 2) growth under high oxygen stress conditions, 3) expression of proteolytically active HtrA oligomers, 4) secretion of HtrA into the supernatant, 5) cell attachment and invasion, and 6) transmigration across polarized epithelial cells. These results establish a genetic complementation system for htrA in C. jejuni, exclude polar effects in the Delta htrA mutants, confirm important HtrA properties, and permit the discovery and dissection of new functions.
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