Cytoplasmic Dynamics Reveals Two Modes of Nucleoid-Dependent Mobility

• Published in: Biophysical journal Vol. 107; no. 11; pp. 2684 - 2692
• Main Authors: Stylianidou, Stella, Kuwada, Nathan J., Wiggins, Paul A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 02.12.2014; Biophysical Society; The Biophysical Society
• Subjects: Biophysics; Cell Biophysics; Cell Cycle; Cytoplasm; Cytoplasm - metabolism; Diffusion; DNA, Bacterial - metabolism; Escherichia coli - cytology; Escherichia coli - growth & development; Escherichia coli - metabolism; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Fluorescence; Membranes; Models, Biological; Molecular biology; Motility; Movement; Protein folding; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA-protein interactions
• ISSN: 0006-3495; 1542-0086
• DOI: 10.1016/j.bpj.2014.10.030

It has been proposed that forces resulting from the physical exclusion of macromolecules from the bacterial nucleoid play a central role in organizing the bacterial cell, yet this proposal has not been quantitatively tested. To investigate this hypothesis, we mapped the generic motion of large protein complexes in the bacterial cytoplasm through quantitative analysis of thousands of complete cell-cycle trajectories of fluorescently tagged ectopic MS2-mRNA complexes. We find the motion of these complexes in the cytoplasm is strongly dependent on their spatial position along the long axis of the cell, and that their dynamics are consistent with a quantitative model that requires only nucleoid exclusion and membrane confinement. This analysis also reveals that the nucleoid increases the mobility of MS2-mRNA complexes, resulting in a fourfold increase in diffusion coefficients between regions of the lowest and highest nucleoid density. These data provide strong quantitative support for two modes of nucleoid action: the widely accepted mechanism of nucleoid exclusion in organizing the cell and a newly proposed mode, in which the nucleoid facilitates rapid motion throughout the cytoplasm.