Team:Osaka/MOTILITY

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MOTILITY

Overview

Under construction

Object

Motile cells, such as Escherichia Coli and Salmonella typhimurium swim by means of flagella. So, we can regard cells as biological paints, we named ColorColi, that can move automatically. Although we can create a lot of artworks (you can see our works at WORKS) by using innate motility of cells, engineering cells motility should expand the application of ColorColi in Art. This year, we aimed to stop the swarming motility of cell for out-put of arithmetic processing by cell-cell communication.(see SIGNAL).


In past iGEM projects, several teams tried to control cell motility (iHKU2008, Imperial2008, Penn State Uni2006). However, their approaches require modified or specified chassis. To control the cell motility more conveniently, more general methods will be needed.


So we made a new part to inhibit flagellar assembly and as a result stop the motility. And in addition, we tested the compatibility of EpsE that work as molecular clutch and stop the motor rotation in B. subtilis for further work.


Design

Bacterial flagellar assembly is proceed by highly sophisticated manner in which gene regulation coordinates with self assembly of motor proteins. [1].To form the flagellar axial structure, "molecular propeller", at the cell exterior, these protein subunits must be translocated across the cell membrane. And this work is carried out by flagellar type III secretion system[2]. By a lot of study, the molecular mechanism of this systems is being elucidated recently.

Currently, the following model for flagellar protein export is suggested.(Fig. 1)N-terminal of segment of a substrate is initially docked with by formation of the FliHx6 complex. And then, ATP hydrolysis induces dissociation of the FliHx6 and successive unfolding and translocation of the substrates is driven by the PMF.

FliH, the regulator of ATPase FliI, is one of soluble components of export apparatus. In the absence of FliI, FliH inhibits the protein translocation [3].

Reference

[1]Fabienne F. V. Chevance and Kelly T. Hughes, “Coordinating assembly of a bacterial macromolecular machine,” Nat Rev Micro 6, no. 6 (June 2008): 455-465.

[2]Tohru Minamino, Katsumi Imada, and Keiichi Namba, “Molecular motors of the bacterial flagella,” Current Opinion in Structural Biology 18, no. 6 (December 2008): 693-701.

[3]Minamino, T. & Namba, K. Distinct roles of the FliI ATPase and proton motive force in bacterial flagellar protein export. Nature 451, 485-488(2008).