An intrinsically disordered linker controlling the formation and the stability of the bacterial flagellar hook

Clive S Barker,Irina V Meshcheryakova,Alla S Kostyukova,Peter L Freddolino,Fadel A Samatey

doi:10.1186/s12915-017-0438-7

Abstract

BackgroundIn a macro-molecular complex, any minor change may prove detrimental. For a supra-molecular nano-machine like the bacterial flagellum, which consists of several distinct parts with specific characteristics, stability is important. During the rotation of the bacterial flagellar motor, which is located in the membrane, the flagella rotate at speeds between 200 and 2000 rpm, depending on the bacterial species. The hook substructure of the bacterial flagellum acts as a universal joint connecting the motor to the flagellar filament. We investigated the formation of the bacterial flagellar hook and its overall stability between the FlgE subunits that make up the hook and attempted to understand how this stability differs between bacteria.ResultsAn intrinsically disordered segment plays an important role for overall hook stability and for its structural cohesion during motor rotation. The length of this linker segment depends on the species of bacteria; for Salmonella enterica and Campylobacter jejuni it is approximately 37 and 54 residues, respectively. Few residues of the linker are conserved and mutating the conserved residues of the linker yields non-flagellated cells. In the case of Campylobacter, which rotates its flagella at a speed much higher than that of Salmonella, shortening the linker leads to a rupture of the hook at its base, decreasing cell motility. Our experiments show that this segment is required for polymerization and stability of the hook, demonstrating a surprising role for a disordered region in one of the most finely tuned and closely studied macromolecular machines.ConclusionsThis study reveals a detailed functional characteristic of an intrinsically disordered segment in the hook protein. This segment evolved to fulfill a specific role in the formation of the hook, and it is at the core of the stability and flexibility of the hook. Its length is important in the case of bacteria with high-speed rotating flagella. Finding a way of disrupting this linker in Campylobacter might help in preventing infections.

Highlights

In a macro-molecular complex, any minor change may prove detrimental
Lys32 of Salmonella FlgE is required for hook assembly In FlgE, the length of the ID-Rod-Stretch is species dependent, while it is conserved in FlgG with a length similar to that of the ID-Rod-Stretch of FlgE from C. jejuni with 54 amino acid residues
In FlgE of S. enterica, the four conserved residues correspond to Thr28, Gly30, Lys32 and Phe38, and the two semiconserved residues correspond to Phe31 and Met41

Summary

Introduction

For a supra-molecular nano-machine like the bacterial flagellum, which consists of several distinct parts with specific characteristics, stability is important. The ID-Rod-Stretch plays an important role in the formation and stability of the hook This linker becomes partly structured upon formation of the hook, where it is located in a pocket surrounded by molecules of FlgE (Fig. 3a, b). To pinpoint which parts of the ID-Rod-Stretch are important for the assembly and function of the hook, we performed targeted mutagenesis of conserved residues in the disordered segment of FlgE from S. enterica. These experiments revealed that this segment is at the center of the stability or flexibility of the hook

Methods

Results

Discussion

Conclusion