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Volume 166, Issue 9

MifS, a DctB family histidine kinase, is a specific regulator of α-ketoglutarate response in PAO1 Free

Abstract

The C5-dicarboxylate α-ketoglutarate (α-KG) is a preferred nutrient source for the opportunistic pathogen . However, very little is known about how detects and responds to α-KG in the environment. Our laboratory has previously shown that the MifS/MifR two-component signal transduction system regulates α-KG assimilation in PAO1. In an effort to better understand how this bacterium detects α-KG, we characterized the MifS sensor histidine kinase. In this study we show that although MifS is a homologue of the C4-dicarboxylate sensor DctB, it specifically responds to the C5-dicarboxylate α-KG. MifS activity increased >10-fold in the presence of α-KG, while the related C5-dicarboxylate glutarate caused only a 2-fold increase in activity. All other dicarboxylates tested did not show any significant effect on MifS activity. Homology modelling of the MifS sensor domain revealed a substrate binding pocket for α-KG. Using protein modelling and mutational analysis, we identified nine residues that are important for α-KG response, including one residue that determines the substrate specificity of MifS. Further, we found that MifS has a novel cytoplasmic linker domain that is required for α-KG response and is probably involved in signal transduction from the sensor domain to the cytoplasmic transmitter domain. Until this study, DctB family histidine kinases were known to only respond to C4-dicarboxylates. Our work shows that MifS is a novel member of the DctB family histidine kinase that specifically responds to α-KG.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): dicarboxylate sensor , MifS , Pseudomonas aeruginosa , sensor histidine kinase , two-component signal transduction systems and α-ketoglutarate
Funding
This study was supported by the:
  • National Institute of Allergy and Infectious Diseases (Award R15GM104880-03)
    • Principle Award Recipient: Christopher T. Nomura
© 2020 The Authors
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2020-06-17
2024-05-10
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MifS, a DctB family histidine kinase, is a specific regulator of α-ketoglutarate response in Pseudomonas aeruginosa PAO1
Microbiology 166, 867 (2020); https://doi.org/10.1099/mic.0.000943
/content/journal/micro/10.1099/mic.0.000943
/content/journal/micro/10.1099/mic.0.000943
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