Bacillus subtilisDuring Feast and Famine: Visualization of the Overall Regulation of Protein Synthesis During Glucose Starvation by Proteome Analysis

Abstract

Dual channel imaging and warping of two-dimensional (2D) protein gels were used to visualize global changes of the gene expression patterns in growing Bacillus subtilis cells during entry into the stationary phase as triggered by glucose exhaustion. The 2D gels only depict single moments during the cells' growth cycle, but a sequential series of overlays obtained at specific points of the growth curve facilitates visualization of the developmental processes at the proteomics scale. During glucose starvation a substantial reprogramming of the protein synthesis pattern was found, with 150 proteins synthesized de novo and cessation of the synthesis of almost 400 proteins. Proteins induced following glucose starvation belong to two main regulation groups: general stress/starvation responses induced by different stresses or starvation stimuli ( B -dependent general stress regulon, stringent response, sporulation), and glucose-starvation-specific responses (drop in glycolysis, utilization of alternative carbon sources, gluconeogenesis). Using the dual channel approach, it was not only possible to identify those regulons or stimulons, but also to follow the fate of each single protein by the three-color code: red, newly induced but not yet accumulated; yellow, synthesized and accumulated; and green, still present, but no longer being synthesized. These green proteins, which represent a substantial part of the protein pool in the nongrowing cell, are not accessible by using DNA arrays. The combination of 2D gel electrophoresis and MALDI TOF mass spectrometry with the dual channel imaging technique provides a new and comprehensive view of the physiology of growing or starving bacterial cell populations, here for the case of the glucosestarvation response.[This is presented as a movie of B. subtilis's growth/glucose-starvation response, available at www.genome.org and also at

The relevance of carbon dioxide metabolism in Streptococcus thermophilus

“…Proteomic analysis of wild-type S. thermophilus and A19(DppcDcarB) mutant grown under different gas atmospheres Proteomic approaches have been widely used to monitor global changes of gene expression patterns during bacterial growth/starvation, or following environmental stresses, and they have provided a comprehensive view in understanding microbial physiology or adaptation to external stimuli (Arena et al, 2006;Bernhardt et al, 2003;Eymann et al, 2004;Salzano et al, 2007). Thus, the effect of different CO 2 Fig.…”

Proteome-wide analysis of the functional roles of bacilysin biosynthesis in Bacillus subtilis

“…Tam et al (2006) complemented the vegetative proteome map of B. subtilis by constructive, another one in response to stress and starvation using the 2D-gel-based approach. In total more than 200 stress or starvation induced proteins have been identified (Antelmann et al, 2000;Bernhardt et al, 2003;Mostertz et al 2004;Tam et al, 2006;Höper et al, 2006). Eighty-three of them were absent in the vegetative proteome map, whereas the remaining stress and starvation induced proteins exhibit a basal expression level already in vegetative growing cells, indicating a basic physiological role in growing and non-growing cells (Wolff et al, 2007).…”

Application of Two-Dimensional Gel Electrophoresis to Microbial Systems

“…The introduction of the highly sensitive technique of 2D-gel electrophoresis into bacterial physiology by Neidhardt and van Bogelen more than 20 years ago, otherwise known as "proteomics" in the mid-90s (Wilkins et al, 1996b), opened a new era in this field of research (Bernhardt et al, 2003;Hecker et al, 2009). …”

“…Recent advances in the application of 2D-gel electrophoresis have been applied to the metabolic system of the Gram positive bacterium, B. subtilis (Bernhardt et al, 2003;Hecker et al, 2009). …”

“…In order to counteract this problem, staining methods have been developed to improve the accuracy and the sensitivity of protein detection and quantification. Nevertheless, several regulatory proteins are present in the cell at very low concentrations, hence, alongside 2D-gel electrophoresis, sensitive high-throughput methods for accurate protein detection and quantification is widely acknowledged (Vemuri et al, 2005) and is currently being implemented (Bernhardt et al, 2003).…”