Dynamics of the bacterial intermediate filament crescentin in vitro and in vivo.

PloS One
Osigwe EsueDenis Wirtz

Abstract

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. 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 microm...Continue Reading

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Methods Mentioned

BETA
rheology
gel filtration
electron microscopy
confocal microscopy
fluorescence microscopy

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