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NAD(P)H-hydrate dehydratase- a metabolic repair enzyme and its role in Bacillus subtilis stress adaptation.

Petrovova M, Tkadlec J, Dvoracek L, Streitova E, Licha I - PLoS ONE (2014)

Bottom Line: The patterns of enzymes for branched amino acid metabolism and cell wall synthesis (L-alanine dehydrogenase, aspartate-semialdehyde dehydrogenase, ketol-acid reductoisomerase) were altered after ethanol stress.We performed the first characterization of a Bacillus subtilis168 knock-out mutant in the yxkO gene that encodes a metabolite repair enzyme.We show that such enzymes could play a significant role in the survival of stressed cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics and Microbiology, Faculty of Science, Charles University, Prague, Czech Republic.

ABSTRACT

Background: One of the strategies for survival stress conditions in bacteria is a regulatory adaptive system called general stress response (GSR), which is dependent on the SigB transcription factor in Bacillus sp. The GSR is one of the largest regulon in Bacillus sp., including about 100 genes; however, most of the genes that show changes in expression during various stresses have not yet been characterized or assigned a biochemical function for the encoded proteins. Previously, we characterized the Bacillus subtilis168 osmosensitive mutant, defective in the yxkO gene (encoding a putative ribokinase), which was recently assigned in vitro as an ADP/ATP-dependent NAD(P)H-hydrate dehydratase and was demonstrated to belong to the SigB operon.

Methods and results: We show the impact of YxkO on the activity of SigB-dependent Pctc promoter and adaptation to osmotic and ethanol stress and potassium limitation respectively. Using a 2DE approach, we compare the proteomes of WT and mutant strains grown under conditions of osmotic and ethanol stress. Both stresses led to changes in the protein level of enzymes that are involved in motility (flagellin), citrate cycle (isocitrate dehydrogenase, malate dehydrogenase), glycolysis (phosphoglycerate kinase), and decomposition of Amadori products (fructosamine-6-phosphate deglycase). Glutamine synthetase revealed a different pattern after osmotic stress. The patterns of enzymes for branched amino acid metabolism and cell wall synthesis (L-alanine dehydrogenase, aspartate-semialdehyde dehydrogenase, ketol-acid reductoisomerase) were altered after ethanol stress.

Conclusion: We performed the first characterization of a Bacillus subtilis168 knock-out mutant in the yxkO gene that encodes a metabolite repair enzyme. We show that such enzymes could play a significant role in the survival of stressed cells.

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Related in: MedlinePlus

Comparative 2DE analysis of WT versus MP2 mutant exposed to osmotic and ethanol stress – stress adaptation and motility.Proteins with protein level profiles that are similar for both stresses. For experimental conditions and data evaluation, see Material and Methods. The picture description is same as for Figure 3.
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pone-0112590-g004: Comparative 2DE analysis of WT versus MP2 mutant exposed to osmotic and ethanol stress – stress adaptation and motility.Proteins with protein level profiles that are similar for both stresses. For experimental conditions and data evaluation, see Material and Methods. The picture description is same as for Figure 3.

Mentions: The results, summarized in Figures 3–6, show that disruption of the yxkO gene caused changes in the level of seven proteins belonging to various metabolic pathways upon both stresses (Figure 3 and Figure 4), and we detected only one protein with a different intensity solely under osmotic stress (Figure 5), three enzymes revealed an altered pattern merely under ethanol stress (Figure 6). Figure S1 illustrates the pattern of the 2DE spot distribution.


NAD(P)H-hydrate dehydratase- a metabolic repair enzyme and its role in Bacillus subtilis stress adaptation.

Petrovova M, Tkadlec J, Dvoracek L, Streitova E, Licha I - PLoS ONE (2014)

Comparative 2DE analysis of WT versus MP2 mutant exposed to osmotic and ethanol stress – stress adaptation and motility.Proteins with protein level profiles that are similar for both stresses. For experimental conditions and data evaluation, see Material and Methods. The picture description is same as for Figure 3.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4231035&req=5

pone-0112590-g004: Comparative 2DE analysis of WT versus MP2 mutant exposed to osmotic and ethanol stress – stress adaptation and motility.Proteins with protein level profiles that are similar for both stresses. For experimental conditions and data evaluation, see Material and Methods. The picture description is same as for Figure 3.
Mentions: The results, summarized in Figures 3–6, show that disruption of the yxkO gene caused changes in the level of seven proteins belonging to various metabolic pathways upon both stresses (Figure 3 and Figure 4), and we detected only one protein with a different intensity solely under osmotic stress (Figure 5), three enzymes revealed an altered pattern merely under ethanol stress (Figure 6). Figure S1 illustrates the pattern of the 2DE spot distribution.

Bottom Line: The patterns of enzymes for branched amino acid metabolism and cell wall synthesis (L-alanine dehydrogenase, aspartate-semialdehyde dehydrogenase, ketol-acid reductoisomerase) were altered after ethanol stress.We performed the first characterization of a Bacillus subtilis168 knock-out mutant in the yxkO gene that encodes a metabolite repair enzyme.We show that such enzymes could play a significant role in the survival of stressed cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics and Microbiology, Faculty of Science, Charles University, Prague, Czech Republic.

ABSTRACT

Background: One of the strategies for survival stress conditions in bacteria is a regulatory adaptive system called general stress response (GSR), which is dependent on the SigB transcription factor in Bacillus sp. The GSR is one of the largest regulon in Bacillus sp., including about 100 genes; however, most of the genes that show changes in expression during various stresses have not yet been characterized or assigned a biochemical function for the encoded proteins. Previously, we characterized the Bacillus subtilis168 osmosensitive mutant, defective in the yxkO gene (encoding a putative ribokinase), which was recently assigned in vitro as an ADP/ATP-dependent NAD(P)H-hydrate dehydratase and was demonstrated to belong to the SigB operon.

Methods and results: We show the impact of YxkO on the activity of SigB-dependent Pctc promoter and adaptation to osmotic and ethanol stress and potassium limitation respectively. Using a 2DE approach, we compare the proteomes of WT and mutant strains grown under conditions of osmotic and ethanol stress. Both stresses led to changes in the protein level of enzymes that are involved in motility (flagellin), citrate cycle (isocitrate dehydrogenase, malate dehydrogenase), glycolysis (phosphoglycerate kinase), and decomposition of Amadori products (fructosamine-6-phosphate deglycase). Glutamine synthetase revealed a different pattern after osmotic stress. The patterns of enzymes for branched amino acid metabolism and cell wall synthesis (L-alanine dehydrogenase, aspartate-semialdehyde dehydrogenase, ketol-acid reductoisomerase) were altered after ethanol stress.

Conclusion: We performed the first characterization of a Bacillus subtilis168 knock-out mutant in the yxkO gene that encodes a metabolite repair enzyme. We show that such enzymes could play a significant role in the survival of stressed cells.

Show MeSH
Related in: MedlinePlus