title: Multimerization of the Virulence-Enhancing Group A Streptococcus Transcription Factor RivR Is Required for Regulatory Activity.
abstract: Group A Streptococcus (GAS) (Streptococcus pyogenes) causes more than 700 million human infections each year. The significant morbidity and mortality rates associated with GAS infections are in part a consequence of the ability of this pathogen to coordinately regulate virulence factor expression during infection. RofA-like protein IV (RivR) is a member of the Mga-like family of transcriptional regulators, and previously we reported that RivR negatively regulates transcription of the hasA and grab virulence factor-encoding genes. Here, we determined that RivR inhibits the ability of GAS to survive and to replicate in human blood. To begin to assess the biochemical basis of RivR activity, we investigated its ability to form multimers, which is a characteristic of Mga-like proteins. We found that RivR forms both dimers and a higher-molecular-mass multimer, which we hypothesize is a tetramer. As cysteine residues are known to contribute to the ability of proteins to dimerize, we created a library of expression plasmids in which each of the four cysteines in RivR was converted to serine. While the C68S RivR protein was essentially unaffected in its ability to dimerize, the C32S and C377S proteins were attenuated, while the C470S protein completely lacked the ability to dimerize. Consistent with dimerization being required for regulatory activity, the C470S RivR protein was unable to repress hasA and grab gene expression in a rivR mutant. Thus, multimer formation is a prerequisite for RivR activity, which supports recent data obtained for other Mga-like family members, suggesting a common regulatory mechanism.
  
  
  
  The modulation of gene transcription is key to the ability of bacterial pathogens to infect hosts to cause disease. Here, we discovered that the group A Streptococcus transcription factor RivR negatively regulates the ability of this pathogen to survive in human blood, and we also began biochemical characterization of this protein. We determined that, in order for RivR to function, it must self-associate, forming both dimers (consisting of two RivR proteins) and higher-order complexes (consisting of more than two RivR proteins). This functional requirement for RivR is shared by other regulators in the same family of proteins, suggesting a common regulatory mechanism. Insight into how these transcription factors function may facilitate the development of novel therapeutic agents targeting their activity.
authors: Center for Molecular Medicine, Department of Microbiology & Immunology, University of Nevada, Reno School of Medicine, Reno, Nevada, USA.
authors: Anupama
authors: A
authors: Ramalinga
authors: Center for Molecular Medicine, Department of Microbiology & Immunology, University of Nevada, Reno School of Medicine, Reno, Nevada, USA.
authors: Jessica L
authors: JL
authors: Danger
authors: Center for Molecular and Translational Human Infectious Diseases Research, The Houston Methodist Research Institute, Houston, Texas, USA.
authors: Nishanth
authors: N
authors: Makthal
authors: Center for Molecular and Translational Human Infectious Diseases Research, The Houston Methodist Research Institute, Houston, Texas, USA.
authors: Muthiah
authors: M
authors: Kumaraswami
authors: Center for Molecular Medicine, Department of Microbiology & Immunology, University of Nevada, Reno School of Medicine, Reno, Nevada, USA psumby@medicine.nevada.edu.
authors: Paul
authors: P
authors: Sumby
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D017354:Point Mutation; D011487:Protein Conformation; D013297:Streptococcus pyogenes / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D014157:Transcription Factors
keywords: Streptococcus pyogenes; multimer; transcription factor; virulence regulation