Protein subassemblies of the Helicobacter pylori Cag type IV secretion system revealed by localization and interaction studies.
Type IV secretion systems are possibly the most versatile protein transport systems in gram-negative bacteria, with substrates ranging from small proteins to large nucleoprotein complexes. In many cases, such as the cag pathogenicity island of Helicobacter pylori, genes encoding components of a type IV secretion system have been identified due to their sequence similarities to prototypical systems such as the VirB system of Agrobacterium tumefaciens. The Cag type IV secretion system contains at least 14 essential apparatus components and several substrate translocation and auxiliary factors, but the functions of most components cannot be inferred from their sequences due to the lack of similarities. In this study, we have performed a comprehensive sequence analysis of all essential or auxiliary Cag components, and we have used antisera raised against a subset of components to determine their subcellular localization. The results suggest that the Cag system contains functional analogues to all VirB components except VirB5. Moreover, we have characterized mutual stabilization effects and performed a comprehensive yeast two-hybrid screening for potential protein-protein interactions. Immunoprecipitation studies resulted in identification of a secretion apparatus subassembly at the outer membrane. Combining these data, we provide a first low-resolution model of the Cag type IV secretion apparatus.
Kutter
Stefan
S
Abteilung Bakteriologie, Max von Pettenkofer Institut für Hygiene und Medizinische Mikrobiologie, Ludwig Maximilians Universität, 80336 München, Germany.
Buhrdorf
Renate
R
Haas
Jürgen
J
Schneider-Brachert
Wulf
W
Haas
Rainer
R
Fischer
Wolfgang
W
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D044404:Genomic Islands / Q000235:genetics; D016480:Helicobacter pylori / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D015151:Immunoblotting; D047468:Immunoprecipitation; D008954:Models, Biological; D011485:Protein Binding; D020798:Two-Hybrid System Techniques
D001426:Bacterial Proteins