Role of methylation in regulating PhoP activity in Salmonella enterica serovar Typhimurium / 上海交通大学学报（医学版）

ABSTRACT

Objective:To investigate the correlation between methylation and the activity of transcriptional regulator PhoP in Salmonella enterica serovar Typhimurium (S. Typhimurium), and screen for the methyltransferase of PhoP. Methods:The methylation level of PhoP in S. Typhimurium under different culture conditions was determined by Western blotting. The transcription levels of phoP and the genes regulated by phoP were examined to screen for the methyltransferase of PhoP after overexpressing methyltransferase candidates predicted by bioinformatics. And then methyltransferase assay was verified in vitro. The transcription levels of phoP and its methyltransferase were determined by real-time PCR in high concentration of Mg2+ or weak acid conditions. Results:As the concentration of Mg2+increased in the medium, the methylation level of PhoP increased, and its methylation level decreased after S. Typhimurium was stimulated by weak acid. In the screening of 9 methyltransferases predicted by bioinformatics, overexpression of STM14_0023 reduced the transcription level of phoP and its downstream genes and the protein level of PhoP in vivo, but knockout of STM14_0023 had no effect on the expression of phoP and its downstream genes. STM14_0023 was capable of methylating PhoP in vitro and could also increase the methylation level of PhoP after overexpression of STM14_0023 in vivo. Under the condition of high concentration of Mg2+ when the expression of phoP was inhibited, the transcriptional level of STM14_0023 was reduced. Conclusion:STM14_0023 is the methyltransferase of PhoP, and methylation inhibits PhoP activity.