J Bacteriol Virol.  2013 Dec;43(4):279-289. 10.4167/jbv.2013.43.4.279.

Comparison of Gene Expression Patterns between Helicobacter pylor 26695 and its Superoxide Dismutase Isogenic Mutant

• Cho MJ
• Lee SG
• Lee KH
• Song JY
• Lee WK
• Baik SC
• Rhee KH
• Youn HS
• Seo JH
• Kang HL

• KMID: 1501996
• DOI: http://doi.org/10.4167/jbv.2013.43.4.279

Abstract

Helicobacter pylori, a causative agent of gastroduodenal diseases, is a Gram-negative microaerophilic bacterium. Although H. pylori locates in the microaerophilic mucous layer, the bacteria would come into contact harmful reactive oxygen species generated by host immune system. It has been reported that H. pylori harbors various defense mechanisms which can protect bacterial cells from oxygen exposure. The change of the gene expression profile of sodB-negative isogenic mutant of H. pylori 26695 was analyzed by high resolution 2-DE followed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and tandem MS and microarray analysis. Eighteen genes and 41 genes were upregulated and downregulated respectively, either transcriptionally or translationally. Expression levels of three genes including trxB, yxjE and ribE that were changed both on a mRNA level and on a protein level were confirmed by RT-PCR analysis. However, change of expression levels of other major antioxidants such as KatA, AhpC and NapA were not detected, which means Sod is regulated by different way from that of KatA and AhpC. Mutant study of other antioxidant proteins may give us better understanding for the regulation of stress response in H. pylori.

Keyword

Helicobacter pylori; Antioxidant; sodB

MeSH Terms

Antioxidants
Bacteria
Defense Mechanisms
Gene Expression*
Helicobacter pylori
Helicobacter*
Immune System
Mass Spectrometry
Microarray Analysis
Oxygen
Reactive Oxygen Species
Ribes
RNA, Messenger
Superoxide Dismutase*
Superoxides*
Transcriptome
Antioxidants
Oxygen
RNA, Messenger
Reactive Oxygen Species
Superoxide Dismutase
Superoxides

Figure

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  Figure 1. Strategy for construction of plasmid for knock-out mutant

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  Figure 2. Localization of SodB spot on the 2-DE gel

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  Figure 3. Growth of H. pylori 26695 and its isogenic mutant at thin layer culture condition

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  Figure 4. Comparative proteome analysis of wild type and sodB isogenic mutant of H. pylori 26695. (A) wild type (B) sodB mutant. Numbers on the gel are spot numbers.

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  Figure 5. Comparison of expression of genes in wild type and sodB mutant. (A) RT-PCR analysis of trxB, yxjE, ribE (B) (C) (D) expression of three genes. Expression levels were measured by either 2-DE analysis or microarray. RT-PCR was used to confirm the results.

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