title: Localization of protein aggregation in Escherichia coli is governed by diffusion and nucleoid macromolecular crowding effect.
abstract: Aggregates of misfolded proteins are a hallmark of many age-related diseases. Recently, they have been linked to aging of Escherichia coli (E. coli) where protein aggregates accumulate at the old pole region of the aging bacterium. Because of the potential of E. coli as a model organism, elucidating aging and protein aggregation in this bacterium may pave the way to significant advances in our global understanding of aging. A first obstacle along this path is to decipher the mechanisms by which protein aggregates are targeted to specific intercellular locations. Here, using an integrated approach based on individual-based modeling, time-lapse fluorescence microscopy and automated image analysis, we show that the movement of aging-related protein aggregates in E. coli is purely diffusive (Brownian). Using single-particle tracking of protein aggregates in live E. coli cells, we estimated the average size and diffusion constant of the aggregates. Our results provide evidence that the aggregates passively diffuse within the cell, with diffusion constants that depend on their size in agreement with the Stokes-Einstein law. However, the aggregate displacements along the cell long axis are confined to a region that roughly corresponds to the nucleoid-free space in the cell pole, thus confirming the importance of increased macromolecular crowding in the nucleoids. We thus used 3D individual-based modeling to show that these three ingredients (diffusion, aggregation and diffusion hindrance in the nucleoids) are sufficient and necessary to reproduce the available experimental data on aggregate localization in the cells. Taken together, our results strongly support the hypothesis that the localization of aging-related protein aggregates in the poles of E. coli results from the coupling of passive diffusion-aggregation with spatially non-homogeneous macromolecular crowding. They further support the importance of "soft" intracellular structuring (based on macromolecular crowding) in diffusion-based protein localization in E. coli.
authors: Institut National de la Santé et de la Recherche Médicale, Unité 1001, Paris, France.
authors: Anne-Sophie
authors: AS
authors: Coquel
authors: Jean-Pascal
authors: JP
authors: Jacob
authors: Mael
authors: M
authors: Primet
authors: Alice
authors: A
authors: Demarez
authors: Mariella
authors: M
authors: Dimiccoli
authors: Thomas
authors: T
authors: Julou
authors: Lionel
authors: L
authors: Moisan
authors: Ariel B
authors: AB
authors: Lindner
authors: Hugues
authors: H
authors: Berry
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D019295:Computational Biology / Q000379:methods; D003198:Computer Simulation; D004058:Diffusion; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D008856:Microscopy, Fluorescence; D015388:Organelles / Q000378:metabolism*; D011485:Protein Binding; D017510:Protein Folding; D021381:Protein Transport
chemical_list: D029968:Escherichia coli Proteins
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