RESUMO
Bacteria use type III secretion injectisomes to inject effector proteins into eukaryotic target cells. Recruitment of effectors to the machinery and the resulting export hierarchy involve the sorting platform. These conserved proteins form pod structures at the cytosolic interface of the injectisome but are also mobile in the cytosol. Photoactivated localization microscopy in Yersinia enterocolitica revealed a direct interaction of the sorting platform proteins SctQ and SctL with effectors in the cytosol of live bacteria. These proteins form larger cytosolic protein complexes involving the ATPase SctN and the membrane connector SctK. The mobility and composition of these mobile pod structures are modulated in the presence of effectors and their chaperones, and upon initiation of secretion, which also increases the number of injectisomes from ~5 to ~18 per bacterium. Our quantitative data support an effector shuttling mechanism, in which sorting platform proteins bind to effectors in the cytosol and deliver the cargo to the export gate at the membrane-bound injectisome.
Assuntos
Sistemas de Secreção Tipo III , Yersinia enterocolitica , Sistemas de Secreção Tipo III/genética , Sistemas de Secreção Tipo III/metabolismo , Yersinia enterocolitica/genética , Yersinia enterocolitica/metabolismo , Citosol/metabolismo , Transporte Proteico , Microscopia de FluorescênciaRESUMO
Requirements engineering of software products for elderly people faces some special challenges to ensure a maximum of user acceptance. Within the scope of a research project, a web-based platform and a mobile app are approached to enable people to live in their own home as long as possible. This paper is about a developed method of interdisciplinary requirements engineering by a team of social scientists in cooperation with computer scientists.