Role of the outermost subdomain of Salmonella flagellin in the filament structure revealed by electron cryomicroscopy.
A mutant strain of Salmonella typhimurium, SJW46, has flagellar filaments supercoiled in the same form as the wild-type strain, SJW1103, and swims normally. However, its flagellar filaments are mechanically unstable and show anomalous behaviors of polymorphism. Flagellin from SJW46 has a large central deletion from Ala204 to Lys292 of SJW1103 flagellin, which has been thought to be located in the outer surface of the filament. Since the filament structure is determined by intersubunit interactions of the terminal regions in the densely packed core of the filament, no serious involvement of the deleted portion was expected in the filament stability and polymorphism. In order to locate the deleted portion and to understand the underlying mechanism of these anomalous characteristics, we carried out structure analysis of the L-type straight filament reconstituted from a mutant flagellin of SJW46 (SJW46S) and compared the structure with that of the SJW1660 filament, which is also the L-type but composed of flagellin with no deletion. The deleted portion was identified as the outermost subdomain, and the structure in the core region showed no appreciable differences. The structure revealed the previously identified folding of flagellin in further detail, and the significance of intersubunit interactions between outer domains, which are present in the SJW1660 filament but absent in the SJW46 filament. This suggests that these contacts have a significant contribution to the filament stability and polymorphic behavior, despite the fact that the contacting surface area occupies only a minor portion of the whole intersubunit interactions.
Mimori-Kiyosue
Y
Y
International Institute for Advanced Research, Matsushita Electric Industrial Co., Ltd, 3-4 Hikaridai, Seika, 619-0237, Japan.
Yamashita
I
I
Fujiyoshi
Y
Y
Yamaguchi
S
S
Namba
K
K
D016935:Caulobacter crescentus / Q000648:ultrastructure; D020285:Cryoelectron Microscopy; D005407:Flagella / Q000235:genetics / Q000648:ultrastructure*; D005408:Flagellin / Q000235:genetics / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted; D008961:Models, Structural; D009154:Mutation; D012486:Salmonella typhimurium / Q000235:genetics / Q000648:ultrastructure*; D017384:Sequence Deletion
D005408:Flagellin