A proteomic view of cell physiology of Bacillus subtilis--bringing the genome sequence to life.
The genome sequence is the "blue-print of life", and the proteomic approach brings this genome sequence to life. Simple model systems are urgently required to "train" this transformation of the genome sequence into life: why not Bacillus subtilis, the model organism for gram-positive bacteria and of functional genomics? By combination of the highly sensitive 2D protein gel electrophoresis with the identification of the protein spots by microsequencing or mass spectrometry we established a 2D protein index of Bacillus subtilis. In order to depict the entire proteome of a B. subtilis cell, alkaline, cell-wall associated, or extracellular proteins were also included. The proteins of this database (see http://microbio2.biologie.uni-greifswald.de:8880/sub2d.htm) were allocated to proteins with house-keeping functions typical of growing cells and to proteins synthesized particularly in non-growing cells. A computer-aided evaluation of the 2D gels loaded with radioactively-labeled proteins from growing or stressed/starved cells proved to be a powerful tool for the analysis of global regulation of the expression of the entire genome. This is shown for the analysis of glycolysis/TCA cycle (house keeping proteins) and for the analysis of the heat stress stimulon. For the heat stress stimulon it is demonstrated how the proteomic approach can be used: (i) to define the structure of a stimulon, (ii) to dissect stimulons into regulons, (iii) to analyze the regulation, structure, and function of unknown regulons, (iv) to define overlapping reguIons or modulons, and finally (v) to explore complex adaptational networks. Furthermore, it will be demonstrated how the "dual channel pattern comparison" or "proteomics signature" (R. VanBogelen) can be used for a comprehensive understanding or prediction of the physiological state of growing or starving cell populations. This is shown for glucose-starved cells. In order to describe the structure and function of gene regulation groups it is generally recommended to complement the proteomics approach with DNA array technologies. Further studies will focus on the analysis of the global regulation of gene expression by the proteomic approach that cannot be addressed by the application of DNA array techniques: the phosphoproteome and its implications in signal transduction; the global control of protein stability; protein targeting and protein secretion.
Hecker
Michael
M
Ernst-Moritz-Arndt-Universität Greifswald, Institut für Mikrobiologie, F.-L.-Jahn-Strasse 15, 17487 Greifswald, Germany. hecker@biologie.uni-greifswald.de
D000222:Adaptation, Physiological / Q000235:genetics / Q000502:physiology; D000818:Animals; D001412:Bacillus subtilis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D006706:Homeostasis / Q000502:physiology; D006801:Humans; D020543:Proteome / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D020539:Sequence Analysis, Protein / Q000379:methods; D015398:Signal Transduction / Q000502:physiology; D013217:Starvation / Q000378:metabolism / Q000503:physiopathology; D013312:Stress, Physiological / Q000378:metabolism / Q000503:physiopathology
D001426:Bacterial Proteins; D020543:Proteome