Kinetics of CagA type IV secretion by Helicobacter pylori and the requirement for substrate unfolding.
Mol Microbiol
; 116(3): 794-807, 2021 09.
Article
em En
| MEDLINE
| ID: mdl-34121254
ABSTRACT
Type IV secretion of effector proteins is an important principle for interaction of human pathogens with their target cells. The corresponding secretion systems may transport a multitude of effector proteins that have to be deployed in the respective spatiotemporal context, or only a single translocated protein, as in the case of the CagA effector protein produced by the human gastric pathogen Helicobacter pylori. For a more detailed analysis of the kinetics and mode of action of CagA type IV secretion by H. pylori, we describe here, a novel, highly sensitive split luciferase-based translocation reporter which can be easily adapted to different end-point or real-time measurements. Using this reporter, we showed that H. pylori cells are able to rapidly inject a limited amount of their CagA supply into cultured gastric epithelial cells. We have further employed the reporter system to address the question whether CagA has to be unfolded prior to translocation by the type IV secretion system. We showed that protein domains co-translocated with CagA as protein fusions are more readily tolerated as substrates than in other secretion systems, but also provide evidence that unfolding of effector proteins is a prerequisite for their transport.
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Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
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Helicobacter pylori
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Transporte Proteico
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Células Epiteliais
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Desdobramento de Proteína
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Sistemas de Secreção Tipo IV
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Antígenos de Bactérias
Limite:
Humans
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article