1. Identification of stringent response-related and potential serological proteins released from Bacillus anthracis overexpressing the RelA/SpoT homolog, Rsh Bant.
- Author
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Kim SK, Park MK, Kim SH, Oh KG, Jung KH, Hong CH, Yoon JW, and Chai YG
- Subjects
- Bacillus anthracis genetics, Bacillus anthracis metabolism, Bacterial Proteins chemistry, Bacterial Proteins genetics, Bacterial Secretion Systems, Electrophoresis, Gel, Two-Dimensional, GTP Pyrophosphokinase chemistry, GTP Pyrophosphokinase genetics, Gene Expression Regulation, Bacterial, Proteomics, Bacillus anthracis enzymology, Bacterial Proteins metabolism, GTP Pyrophosphokinase metabolism
- Abstract
RelA and SpoT synthesize ppGpp, a key effector molecule that facilitates the adaptation of bacteria to nutrient starvation and other stresses, known as the stringent response. To investigate the role of Rsh Bant , a putative RelA/SpoT homolog (encoded by BAS4302) in Bacillus anthracis, we examined the alteration of the secretome profiles after the overexpression of a functional His-Rsh Bant protein in the B. anthracis strain Sterne at the stationary growth phase. In the ppGpp-deficient E. coli mutant strain CF1693, overexpression of Rsh Bant restored a ppGpp-dependent growth defect on minimal glucose media. The secretome profiles obtained using a two-dimensional electrophoresis (2-DE) analysis were altered by overexpression of Rsh Bant in B. anthracis. Among the 66 protein spots differentially expressed >1.5-fold, the 29 proteins were abundant for further identification using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Functional categorization of those proteins implicated their involvement in various biological activities. Taken together, our results imply that overexpression of a functional His-Rsh Bant can lead to the increased levels of intracellular ppGpp in B. anthracis, resulting in the significant changes in its secretome profiling. The stringent response-controlled proteins identified are likely useful as potential targets for serodiagnostic applications.
- Published
- 2014
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