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Proteomic analysis of the function of sigma factor σ54 in Helicobacter pylori survival with nutrition deficiency stress in vitro.

Sun Y, Liu S, Li W, Shan Y, Li X, Lu X, Li Y, Guo Q, Zhou Y, Jia J - PLoS ONE (2013)

Bottom Line: Our results show that the expression of σ(54) (rpoN) is significantly induced in the stationary phase (nutrition deficiency) and the rpoN mutant showed a significantly lower viability than wild-type H. pylori in the late stationary phase.We used comparative proteomics to analyze the protein differentiation between wild-type and rpoN mutant during the stationary phase.With depleted nutrients, σ(54) can slow the process of proliferation by negatively regulating genes involved in energy metabolism and biosynthesis and enhance stress-resistant ability by positively regulating genes involved in protein fate and redox reaction.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Key Laboratory for Experimental Teratology of Chinese Ministry of Education, School of Medicine, Shandong University, Jinan, Shandong, China.

ABSTRACT
H. pylori can survive under a nutrition-deficient environment. During infection and transmission, H. pylori is confronted with nutrient limitation and the bacterium requires rapid alteration in gene expression for survival under stress conditions. However, the mechanism underlining this regulation remains unknown. A previous study showed that σ(54) is an important regulation factor for H. pylori survival in the nutrition-deficient environment. Our results show that the expression of σ(54) (rpoN) is significantly induced in the stationary phase (nutrition deficiency) and the rpoN mutant showed a significantly lower viability than wild-type H. pylori in the late stationary phase. Thus, σ(54) is involved in H. pylori survival during nutrient limitation. We used comparative proteomics to analyze the protein differentiation between wild-type and rpoN mutant during the stationary phase. With depleted nutrients, σ(54) can slow the process of proliferation by negatively regulating genes involved in energy metabolism and biosynthesis and enhance stress-resistant ability by positively regulating genes involved in protein fate and redox reaction. Especially, NapA positively regulated by σ(54) plays an important function in H. pylori survival both in the stationary phase and in water, and the latter situation would be beneficial for bacterial in vitro transmission. Our investigations give new light on the adaptive regulation of H. pylori under stress conditions.

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mRNA expression confirms the protein expression of most genes from 2-DE maps.mRNA and protein levels of the wild-type (WT) and ΔrpoN involved in energy metabolism (A), antioxidation (B), biosynthesis (C) and protein fate (D). (a) Quantification of total protein spot volume ratio from 2-DE analysis; (b) RT-PCR analysis of mRNA levels. Box-and-whisker plots represent the median value with 50% of all data falling within the box. The “whiskers” extend to the 5th and 95th percentiles. Signals were normalized to 16S rRNA levels. Data are mean ± SEM of replicated experiments. *P <0.05, **P <0.01, ***P <0.001.
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pone-0072920-g003: mRNA expression confirms the protein expression of most genes from 2-DE maps.mRNA and protein levels of the wild-type (WT) and ΔrpoN involved in energy metabolism (A), antioxidation (B), biosynthesis (C) and protein fate (D). (a) Quantification of total protein spot volume ratio from 2-DE analysis; (b) RT-PCR analysis of mRNA levels. Box-and-whisker plots represent the median value with 50% of all data falling within the box. The “whiskers” extend to the 5th and 95th percentiles. Signals were normalized to 16S rRNA levels. Data are mean ± SEM of replicated experiments. *P <0.05, **P <0.01, ***P <0.001.

Mentions: We used proteomic analysis to compare the protein expression profiles of H. pylori 26695 and its rpoN mutant in the early stationary phase to determine the mechanism by which σ54 regulates H. pylori survival. Compared with the proteome of wild-type H. pylori, 27 protein spots were differentially expressed in the rpoN mutant (Figure 2A; Table 2). The functions of these proteins were involved in energy metabolism, biosynthesis, redox reaction, protein fate, and so on (Table 2; Figure 2B and C). The differential expression of these proteins was further confirmed at the mRNA level by quantitative real-time PCR (Figure 3).


Proteomic analysis of the function of sigma factor σ54 in Helicobacter pylori survival with nutrition deficiency stress in vitro.

Sun Y, Liu S, Li W, Shan Y, Li X, Lu X, Li Y, Guo Q, Zhou Y, Jia J - PLoS ONE (2013)

mRNA expression confirms the protein expression of most genes from 2-DE maps.mRNA and protein levels of the wild-type (WT) and ΔrpoN involved in energy metabolism (A), antioxidation (B), biosynthesis (C) and protein fate (D). (a) Quantification of total protein spot volume ratio from 2-DE analysis; (b) RT-PCR analysis of mRNA levels. Box-and-whisker plots represent the median value with 50% of all data falling within the box. The “whiskers” extend to the 5th and 95th percentiles. Signals were normalized to 16S rRNA levels. Data are mean ± SEM of replicated experiments. *P <0.05, **P <0.01, ***P <0.001.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3755968&req=5

pone-0072920-g003: mRNA expression confirms the protein expression of most genes from 2-DE maps.mRNA and protein levels of the wild-type (WT) and ΔrpoN involved in energy metabolism (A), antioxidation (B), biosynthesis (C) and protein fate (D). (a) Quantification of total protein spot volume ratio from 2-DE analysis; (b) RT-PCR analysis of mRNA levels. Box-and-whisker plots represent the median value with 50% of all data falling within the box. The “whiskers” extend to the 5th and 95th percentiles. Signals were normalized to 16S rRNA levels. Data are mean ± SEM of replicated experiments. *P <0.05, **P <0.01, ***P <0.001.
Mentions: We used proteomic analysis to compare the protein expression profiles of H. pylori 26695 and its rpoN mutant in the early stationary phase to determine the mechanism by which σ54 regulates H. pylori survival. Compared with the proteome of wild-type H. pylori, 27 protein spots were differentially expressed in the rpoN mutant (Figure 2A; Table 2). The functions of these proteins were involved in energy metabolism, biosynthesis, redox reaction, protein fate, and so on (Table 2; Figure 2B and C). The differential expression of these proteins was further confirmed at the mRNA level by quantitative real-time PCR (Figure 3).

Bottom Line: Our results show that the expression of σ(54) (rpoN) is significantly induced in the stationary phase (nutrition deficiency) and the rpoN mutant showed a significantly lower viability than wild-type H. pylori in the late stationary phase.We used comparative proteomics to analyze the protein differentiation between wild-type and rpoN mutant during the stationary phase.With depleted nutrients, σ(54) can slow the process of proliferation by negatively regulating genes involved in energy metabolism and biosynthesis and enhance stress-resistant ability by positively regulating genes involved in protein fate and redox reaction.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Key Laboratory for Experimental Teratology of Chinese Ministry of Education, School of Medicine, Shandong University, Jinan, Shandong, China.

ABSTRACT
H. pylori can survive under a nutrition-deficient environment. During infection and transmission, H. pylori is confronted with nutrient limitation and the bacterium requires rapid alteration in gene expression for survival under stress conditions. However, the mechanism underlining this regulation remains unknown. A previous study showed that σ(54) is an important regulation factor for H. pylori survival in the nutrition-deficient environment. Our results show that the expression of σ(54) (rpoN) is significantly induced in the stationary phase (nutrition deficiency) and the rpoN mutant showed a significantly lower viability than wild-type H. pylori in the late stationary phase. Thus, σ(54) is involved in H. pylori survival during nutrient limitation. We used comparative proteomics to analyze the protein differentiation between wild-type and rpoN mutant during the stationary phase. With depleted nutrients, σ(54) can slow the process of proliferation by negatively regulating genes involved in energy metabolism and biosynthesis and enhance stress-resistant ability by positively regulating genes involved in protein fate and redox reaction. Especially, NapA positively regulated by σ(54) plays an important function in H. pylori survival both in the stationary phase and in water, and the latter situation would be beneficial for bacterial in vitro transmission. Our investigations give new light on the adaptive regulation of H. pylori under stress conditions.

Show MeSH
Related in: MedlinePlus