title: Characterization of the Group A Streptococcus Mga virulence regulator reveals a role for the C-terminal region in oligomerization and transcriptional activation.
abstract: The Group A Streptococcus (GAS) is a strict human pathogen that causes a broad spectrum of illnesses. One of the key regulators of virulence in GAS is the transcriptional activator Mga, which co-ordinates the early stages of infection. Although the targets of Mga have been well characterized, basic biochemical analyses have been limited due to difficulties in obtaining purified protein. In this study, high-level purification of soluble Mga was achieved, enabling the first detailed characterization of the protein. Fluorescence titrations coupled with filter-binding assays indicate that Mga binds cognate DNA with nanomolar affinity. Gel filtration analyses, analytical ultracentrifugation and co-immunoprecipitation experiments demonstrate that Mga forms oligomers in solution.Moreover, the ability of the protein to oligomerize in solution was found to correlate with transcriptional activation; DNA binding appears to be necessary but insufficient for full activity. Truncation analyses reveal that the uncharacterized C-terminal region of Mga, possessing similarity to phosphotransferase system EIIB proteins, plays a critical role in oligomerization and in vivo activity. Mga from a divergent serotype was found to behave similarly, suggesting that this study describes a general mechanism for Mga regulation of target virulence genes within GAS and provides insight into related regulators in other Gram-positive pathogens.
authors.affiliation: Department of Cell Biology & Molecular Genetics andMaryland Pathogen Research Institute, University of Maryland, College Park, MD 20742, USA.
authors.forename: Elise R
authors.initials: ER
authors.lastname: Hondorp
authors.forename: Sherry C
authors.initials: SC
authors.lastname: Hou
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Hempstead
authors.forename: Lara L
authors.initials: LL
authors.lastname: Hause
authors.forename: Dorothy M
authors.initials: DM
authors.lastname: Beckett
authors.forename: Kevin S
authors.initials: KS
authors.lastname: McIver
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D055503:Protein Multimerization; D013297:Streptococcus pyogenes / Q000235:genetics* / Q000472:pathogenicity; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D014157:Transcription Factors; C068267:mry protein, Streptococcus pyogenes