Poster Presentation Lancefield International Symposium for Streptococci and Streptococcal Diseases 2025

Exploring the structural basis for CovR dysregulation in Australian M1UK Group A Streptococcus   (#234)

Jarrad Pritchard 1 , Emma-Jayne Proctor 1 , Stephan Brouwer 2 , Jiawa Wang 1 , Nikolas Johnston 1 , Andrew Hayes 3 , Haibo Yu 1 , Jai Tree 4 , Mark Davies 3 , Mark Walker 2 , Ronald Sluyter 1 , Martina Sanderson-Smith 1
  1. School of Science and Molecular Horizons, University of Wollongong, Wollongong, NSW, Australia
  2. Institute for Molecular Bioscience, University of Queensland, Brisbane, QLD, Australia
  3. The Peter Doherty Institute for Infection and Immunity and Department of Microbiology and Immunology, University of Melbourne, Melbourne, VIC, Australia
  4. School of Biotechnology & Biomolecular Sciences, The University of New South Wales, Sydney, NSW, Australia

A worldwide surge in invasive Group A Streptococcal (GAS) infection has now been linked to the emergence of the novel M1UK lineage. Genomic screening of Australian invasive M1UK clinical isolates identified a mutation of A111V in CovR, a component of the CovRS two-component regulatory system. Normally, CovR (transcriptional regulator) dimerises upon phosphorylation by CovS (histidine kinase), driving transcriptional regulation of up to 15% of the GAS genome. Mutations in CovRS, or CovRS switching, have been implicated in the upregulation of several virulence factors in other GAS lineages (e.g. M1T1; a predecessor lineage to M1UK) resulting in a hypervirulent phenotype. Further genomic screening of clinical M1UK GAS isolates identified novel covRS mutations and revealed a comparable CovRS switching rate as observed for M1T1. In vitro, M1UK containing CovRA111V exhibit a hypervirulent phenotype similar to M1T1, characterised by decreased cysteine protease expression and increased expression of haemolytic toxins and hyaluronic acid capsule, promoting immune evasion. While the effect of CovRS switching on GAS virulence is well established, less is known about how CovR mutations affect protein function. We demonstrate an inability of recombinant CovRA111V to dimerise in a phosphorylation-dependent manner. In parallel, molecular dynamics simulations suggest that the A111V mutation specifically destabilises the interface between CovR monomers that facilitate dimerisation, inhibiting CovR dimer-mediated transcriptional repression of virulence genes and promoting a hypervirulent phenotype. Together, these data demonstrate further evidence of covRS switching in M1UK and provide insight into the structural basis for dysregulation of CovR during infection.