RRC ID 34745
著者 Vischer NO, Verheul J, Postma M, van den Berg van Saparoea B, Galli E, Natale P, Gerdes K, Luirink J, Vollmer W, Vicente M, den Blaauwen T.
タイトル Cell age dependent concentration of Escherichia coli divisome proteins analyzed with ImageJ and ObjectJ.
ジャーナル Front Microbiol
Abstract The rod-shaped Gram-negative bacterium Escherichia coli 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°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.
巻・号 6
ページ 586
公開日 2015-1-1
DOI 10.3389/fmicb.2015.00586
PMID 26124755
PMC PMC4462998
IF 4.236
引用数 40
WOS 分野 MICROBIOLOGY
リソース情報
原核生物(大腸菌) ME9062(BW25113) JW0145-KC(mrcB) JW3359-KC(mrcA)