Dynamics of the bacterial intermediate filament crescentin in vitro and in vivo.
BACKGROUND
Crescentin, the recently discovered bacterial intermediate filament protein, organizes into an extended filamentous structure that spans the length of the bacterium Caulobacter crescentus and plays a critical role in defining its curvature. The mechanism by which crescentin mediates cell curvature and whether crescentin filamentous structures are dynamic and/or polar are not fully understood.


RESULTS
Using light microscopy, electron microscopy and quantitative rheology, we investigated the mechanics and dynamics of crescentin structures. Live-cell microscopy reveals that crescentin forms structures in vivo that undergo slow remodeling. The exchange of subunits between these structures and a pool of unassembled subunits is slow during the life cycle of the cell however; in vitro assembly and gelation of C. crescentus crescentin structures are rapid. Moreover, crescentin forms filamentous structures that are elastic, solid-like, and, like other intermediate filaments, can recover a significant portion of their network elasticity after shear. The assembly efficiency of crescentin is largely unaffected by monovalent cations (K(+), Na(+)), but is enhanced by divalent cations (Mg(2+), Ca(2+)), suggesting that the assembly kinetics and micromechanics of crescentin depend on the valence of the ions present in solution.


CONCLUSIONS
These results indicate that crescentin forms filamentous structures that are elastic, labile, and stiff, and that their low dissociation rate from established structures controls the slow remodeling of crescentin in C. crescentus.
Department of Pharmaceutical Development, Genentech, South San Francisco, California, United States of America. esue.osigwe@gene.com
Osigwe
O
Esue
Laura
L
Rupprecht
Sean X
SX
Sun
Denis
D
Wirtz
D001426:Bacterial Proteins / Q000378:metabolism / Q000502:physiology*; D002413:Cations, Divalent; D016934:Caulobacter / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D012212:Rheology
D001426:Bacterial Proteins; D002413:Cations, Divalent