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Cell age dependent concentration of Escherichia coli divisome proteins analyzed with ImageJ and ObjectJ

Lookup NU author(s): Elisa Galli, Professor Kenn Gerdes, Professor Waldemar Vollmer

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Abstract

The rod-shaped Gram-negative bacterium Escherichia colt multiplies by elongation followed by binary fission. Longitudinal growth of the cell envelope and synthesis of the new poles are organized by two protein complexes called elongasome and divisome, respectively. We have analyzed the spatio-temporal localization patterns of many of these morphogenetic proteins by immunolabeling the wild type strain MC4100 grown to steady state in minimal glucose medium at 28 degrees C. This allowed the direct comparison of morphogenetic protein localization patterns as a function of cell age as imaged by phase contrast and fluorescence wide field microscopy. Under steady state conditions the age distribution of the cells is constant and is directly correlated to cell length. To quantify cell size and protein localization parameters in 1000s of labeled cells, we developed 'Coli-Inspector,' which is a project running under ImageJ with the plugin 'ObjectJ.' ObjectJ organizes image-analysis tasks using an integrated approach with the flexibility to produce different output formats from existing markers such as intensity data and geometrical parameters. ObjectJ supports the combination of automatic and interactive methods giving the user complete control over the method of image analysis and data collection, with visual inspection tools for quick elimination of artifacts. Coli-inspector was used to sort the cells according to division cycle cell age and to analyze the spatio-temporal localization pattern of each protein. A unique dataset has been created on the concentration and position of the proteins during the cell cycle. We show for the first time that a subset of morphogenetic proteins have a constant cellular concentration during the cell division cycle whereas another set exhibits a cell division cycle dependent concentration variation. Using the number of proteins present at midcell, the stoichiometry of the divisome is discussed.


Publication metadata

Author(s): Vischer NOE, Verheul J, Postma M, van Saparoea BV, Galli E, Natale P, Gerdes K, Luirink J, Vollmer W, Vicente M, den Blaauwen T

Publication type: Article

Publication status: Published

Journal: Frontiers in Microbiology

Year: 2015

Volume: 6

Online publication date: 11/06/2015

Acceptance date: 28/05/2015

ISSN (electronic): 1664-302X

Publisher: Frontiers Research Foundation

URL: http://dx.doi.org/10.3389/fmicb.2015.00586

DOI: 10.3389/fmicb.2015.00586


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