TY - JOUR
T1 - Staphylococcus aureus PhoU Homologs Regulate Persister Formation and Virulence
AU - Shang, Yongpeng
AU - Wang, Xiaofei
AU - Chen, Zhong
AU - Lyu, Zhihui
AU - Lin, Zhiwei
AU - Zheng, Jinxin
AU - Wu, Yang
AU - Deng, Qiwen
AU - Yu, Zhijian
AU - Zhang, Ying
AU - Qu, Di
N1 - Funding Information:
We gratefully acknowledge Dr. Tao Xu (Department of Infectious Diseases, Huashan Hospital, Fudan University, Shanghai, China) for his suggestions on the gene knockout. Funding. This work was supported by the National Natural Science Foundation of China (81871622, 81271791, 81571955, 81671982, and 81991532), the National Major Scientific and Technological Projects in “Infectious Disease” (2018ZX10734401), “Development of Key Technologies and Drugs for the Prevention of Drug-Resistant Bacteria” (2019ZX09721001), and “Research on Biosafety Technology of High-Level Biosafety Laboratory and Important Pathogen Laboratory” (2018ZX10734401-04), and the Sanming Project of Medicine in Shenzhen (SMGC201705029).
Funding Information:
This work was supported by the National Natural Science Foundation of China (81871622, 81271791, 81571955, 81671982, and 81991532), the National Major Scientific and Technological Projects in “Infectious Disease” (2018ZX10734401), “Development of Key Technologies and Drugs for the Prevention of Drug-Resistant Bacteria” (2019ZX09721001), and “Research on Biosafety Technology of High-Level Biosafety Laboratory and Important Pathogen Laboratory” (2018ZX10734401-04), and the Sanming Project of Medicine in Shenzhen (SMGC201705029).
Publisher Copyright:
© Copyright © 2020 Shang, Wang, Chen, Lyu, Lin, Zheng, Wu, Deng, Yu, Zhang and Qu.
PY - 2020/5/26
Y1 - 2020/5/26
N2 - PhoU homologs are one of the determinant factors in the regulation of persister formation and phosphate metabolism in many bacterial species; however, the functions of PhoU homologs exhibit species-specific characteristics. The pathogenesis of Staphylococcus aureus is closely correlated with persister formation and virulence factors. The functions of two PhoU homologs, PhoU1 and PhoU2, in S. aureus are unclear yet. In this study, single- and double-deletion mutants of phoU1 and phoU2 were generated in strain USA500 2395. The ΔphoU1 or ΔphoU2 mutants displayed a change in persister formation and virulence compared to the parent strain; the persisters to vancomycin and levofloxacin were decreased at least 1,000-fold, and the number of intracellular bacteria surviving in the A549 cells for 24 h decreased to 82 or 85%. The α-hemolysin expression and activity were increased in the ΔphoU2 mutants. Transcriptome analysis revealed that 573 or 285 genes were differentially expressed by at least 2.0-fold in the ΔphoU1 or ΔphoU2 mutant vs. the wild type. Genes involved in carbon and pyruvate metabolism were up-regulated, and virulence genes and virulence regulatory genes were down-regulated, including type VII secretion system, serine protease, leukocidin, global regulator (sarA, rot), and the two-component signal transduction system (saeS). Correspondingly, the deletion of the phoU1 or phoU2 resulted in increased levels of intracellular pyruvate and ATP. Deletion of the phoU2, but not the phoU1, resulted in the up-regulation of inorganic phosphate transport genes and increased levels of intracellular inorganic polyphosphate. In conclusion, both PhoU1 and PhoU2 in S. aureus regulate virulence by the down-regulation of multiple virulence factors (type VII secretion system, serine protease, and leucocidin) and the persister generation by hyperactive carbon metabolism accompanied by increasing intracellular ATP. The results in S. aureus are different from what we have previously found in Staphylococcus epidermis, where only PhoU2 regulates biofilm and persister formation. The different functions of PhoU homologs between the two species of Staphylococcus warrant further investigation.
AB - PhoU homologs are one of the determinant factors in the regulation of persister formation and phosphate metabolism in many bacterial species; however, the functions of PhoU homologs exhibit species-specific characteristics. The pathogenesis of Staphylococcus aureus is closely correlated with persister formation and virulence factors. The functions of two PhoU homologs, PhoU1 and PhoU2, in S. aureus are unclear yet. In this study, single- and double-deletion mutants of phoU1 and phoU2 were generated in strain USA500 2395. The ΔphoU1 or ΔphoU2 mutants displayed a change in persister formation and virulence compared to the parent strain; the persisters to vancomycin and levofloxacin were decreased at least 1,000-fold, and the number of intracellular bacteria surviving in the A549 cells for 24 h decreased to 82 or 85%. The α-hemolysin expression and activity were increased in the ΔphoU2 mutants. Transcriptome analysis revealed that 573 or 285 genes were differentially expressed by at least 2.0-fold in the ΔphoU1 or ΔphoU2 mutant vs. the wild type. Genes involved in carbon and pyruvate metabolism were up-regulated, and virulence genes and virulence regulatory genes were down-regulated, including type VII secretion system, serine protease, leukocidin, global regulator (sarA, rot), and the two-component signal transduction system (saeS). Correspondingly, the deletion of the phoU1 or phoU2 resulted in increased levels of intracellular pyruvate and ATP. Deletion of the phoU2, but not the phoU1, resulted in the up-regulation of inorganic phosphate transport genes and increased levels of intracellular inorganic polyphosphate. In conclusion, both PhoU1 and PhoU2 in S. aureus regulate virulence by the down-regulation of multiple virulence factors (type VII secretion system, serine protease, and leucocidin) and the persister generation by hyperactive carbon metabolism accompanied by increasing intracellular ATP. The results in S. aureus are different from what we have previously found in Staphylococcus epidermis, where only PhoU2 regulates biofilm and persister formation. The different functions of PhoU homologs between the two species of Staphylococcus warrant further investigation.
KW - ATP
KW - PhoU homolog
KW - Staphylococcus aureus
KW - persisters
KW - phosphate metabolism
KW - virulence
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U2 - 10.3389/fmicb.2020.00865
DO - 10.3389/fmicb.2020.00865
M3 - Article
C2 - 32670206
AN - SCOPUS:85086248096
SN - 1664-302X
VL - 11
JO - Frontiers in Microbiology
JF - Frontiers in Microbiology
M1 - 865
ER -