title: Extracellular secretion of protease HtrA from Campylobacter jejuni is highly efficient and independent of its protease activity and flagellum.
abstract: The serine protease HtrA of C. jejuni has been identified as a novel secreted virulence factor which opens cell-to-cell junctions by cleaving E-cadherin. Efficient C. jejuni transmigration across polarized human epithelial cells requires the intact flagellum and HtrA; however, the mechanism of HtrA secretion into the supernatant is unknown. Here we show that HtrA secretion is highly efficient and does not require its proteolytic activity because the protease-inactive S197A mutant is secreted like wild-type HtrA. In addition, the flagellar mutants ΔflaA/B, ΔfliI, ΔflgH, ΔflhA, ΔflhB, and ΔflgS were also able to secrete HtrA in high amounts, while they were strongly attenuated in secreting the well-known invasion antigen CiaB. We also tested several culture media and cell lines of different origin such as human, mouse, hamster, dog, and chicken for their ability to influence HtrA secretion. Interestingly, HtrA was effectively secreted in the presence of most but not all cell lines and media, albeit at different levels, but secretion was significantly higher when fetal calf serum (FCS) was added. These results demonstrate that HtrA secretion by Campylobacter proceeds independent of HtrA's protease activity, the flagellum and origin of cell lines, but can be strongly enhanced by molecular compound(s) present in FCS.
authors.forename: Manja
authors.initials: M
authors.lastname: Boehm
authors.forename: Ingrid
authors.initials: I
authors.lastname: Haenel
authors.forename: Benjamin
authors.initials: B
authors.lastname: Hoy
authors.forename: Lone
authors.initials: L
authors.lastname: Brøndsted
authors.forename: Todd G
authors.initials: TG
authors.lastname: Smith
authors.forename: Timothy
authors.initials: T
authors.lastname: Hoover
authors.forename: Silja
authors.initials: S
authors.lastname: Wessler
authors.forename: Nicole
authors.initials: N
authors.lastname: Tegtmeyer
keywords: E-cadherin; HtrA; cellular invasion; flagellum; molecular pathogenesis; secretion; signaling; virulence