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Functional analysis of the sortase YhcS in Bacillus subtilis
Hamidreza Fasehee1, Helga Westers, Albert Bolhuis
1Department of Molecular Genetics, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Proteomics
|July 30, 2011
Summary
Bacillus subtilis possesses a functional sortase, YhcS, which anchors proteins like YfkN to the cell wall. This sortase activity is crucial for retaining specific proteins, preventing their release during phosphate starvation.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- Sortases are essential enzymes in Gram-positive bacteria, catalyzing protein anchoring to the cell wall.
- Bacillus subtilis is a key host for protein production, with two potential sortases, YhcS and YwpE.
Purpose of the Study:
- To investigate the sortase function of YhcS and YwpE in Bacillus subtilis.
- To identify and characterize sortase substrates and their anchoring mechanisms.
Main Methods:
- Proteomics analysis to identify released proteins in sortase-mutant cells.
- Gene expression analysis and protein fusion studies using reporter proteins (e.g., chitinase ChiS).
Main Results:
- Sortase mutants showed increased release of the putative substrate YfkN during phosphate starvation.
- YfkN requires YhcS sortase activity for cell wall retention.
- A functional cell-wall-anchoring motif from YhcR was demonstrated when fused to chitinase ChiS, enabling cell wall display via YhcS.
Conclusions:
- Bacillus subtilis possesses a functional sortase (YhcS) capable of anchoring proteins to the cell wall.
- This sortase activity plays a role in regulating protein localization, particularly under nutrient stress.
- The study provides insights into protein secretion and cell wall modification in B. subtilis.
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