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Volume 166, Issue 4

Control of acetyl phosphate-dependent phosphorylation of the response regulator CiaR by acetate kinase in Free

Abstract

The two-component regulatory system CiaRH of affects a large variety of physiological processes including ß-lactam resistance, competence development, maintenance of cell integrity, bacteriocin production, but also host colonization and virulence. The response regulator CiaR is active under a wide variety of conditions and the cognate CiaH kinase is not always needed to maintain CiaR activity. Using tetracycline-controlled expression of and variants, acetyl phosphate was identified as the alternative source of CiaR phosphorylation in the absence of CiaH. Concomitant inactivation of and the acetate kinase gene led to very high levels of CiaR-mediated promoter activation. Strong transcriptional activation was accompanied by a high phosphorylation status of CiaR as determined by Phos-tag gel electrophoresis of cell extracts. Furthermore, AckA acted negatively upon acetyl phosphate-dependent phosphorylation of CiaR. Experiments using the two-hybrid system based on adenylate cyclase reconstitution indicated binding of AckA to CiaR and therefore direct regulation. Subsequent CiaR phosphorylation experiments confirmed observations. Purified AckA was able to inhibit acetyl phosphate-dependent phosphorylation. Inhibition required the presence of ADP. AckA-mediated regulation of CiaR phosphorylation is the first example for a regulatory connection of acetate kinase to a response regulator besides controlling acetyl phosphate levels. It will be interesting to see if this novel regulation applies to other response regulators in or even in other organisms.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): acetate kinase AckA , acetyl phosphate-dependent phosphorylation , in vitrophosphorylation of CiaR , Streptococcus pneumoniae and two-component regulatory system CiaRH; CiaR mutant proteins
Funding
This study was supported by the:
  • Deutsche Forschungsgemeinschaft (Award BR947-8-1)
    • Principle Award Recipient: Reinhold Brückner
© 2020 The Authors
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2020-02-19
2024-04-19
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Control of acetyl phosphate-dependent phosphorylation of the response regulator CiaR by acetate kinase in Streptococcus pneumoniae
Microbiology 166, 411 (2020); https://doi.org/10.1099/mic.0.000894
/content/journal/micro/10.1099/mic.0.000894
/content/journal/micro/10.1099/mic.0.000894
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