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Identification of Prx binding partners from Streptococcus pyogenes lysates

ABSTRACT: The small prophage protein paratox (Prx) from Streptococcus pyogenes strain MGAS315 was used as bait in pull-down assays with bacterial lysates to discover potential binding partners. Bacterial lysates analyzed included MGAS315 stimulated with the quorum sensing pheromone XIP-- that induces natural competence, and MGAS315 stimulated with mitomycin C which induces bacteriophage protein expression.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Streptococcus Pyogenes Mgas315 (ncbitaxon:198466)

SUBMITTER: Gerd Prehna

PROVIDER: MSV000095491 | MassIVE | Wed Jul 31 06:42:00 BST 2024

REPOSITORIES: MassIVE

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Project description:Streptococcus pyogenes is a major causative agent of tonsillitis or pharyngitis in children, which can lead to more invasive infections and noninfectious sequellae. S. pyogenes can persist in tonsils, while one-third of children treated with antibiotics continue to shed streptococci and have recurrent infections. Mouse nasal-associated lymphoid tissue (NALT) is functionally analogous to human oropharangeal lymphoid tissues. The innate immune responses of naïve cells from a mucosal site to S. pyogenes is not well described; therefore, we infected C57BL/6 mice intranasally with 108 CFU S. pyogenes. Transcriptional responses by NALT after S. pyogenes infection were analyzed by Affymetrix microarray and quantitative RT-PCR. Wild-type S. pyogenes induces transcription of both type I and IFN-gamma-responsive genes, pro-inflammatory genes, and acute phase response plasma proteins within 24h after infection. Invasion of NALT and the induction of the interferon response were not dependent on expression of anti-phagocytic M1 protein. However, infection with an attenuated, less invasive mutant indicated that a robust innate response by NALT is significantly influenced by intra-NALT bacterial load. Granulocytic populations of NALT, cervical lymph nodes and spleen were discriminated by characteristic surface and intracellular markers. Intranasal infection induces systemic release of neutrophils and a substantial influx of neutrophils into NALT at 24h, which decline by 48h after infection. Macrophages do not significantly increase in S. pyogenes-infected NALT. Intranasal infection of IFN-gamma -/- (GKO) C57BL/6 mice did not lead to systemic dissemination of wild type S. pyogenes, despite reduced expression of IFN-gamma-responsive mRNAs in NALT. Infected GKO mice had an unregulated influx of neutrophils into NALT compared to immunocompetant mice and mice treated with an anti-IFN-gamma antibody more rapidly cleared S. pyogenes from NALT. Thus, IFN-gamma-induced responses have a suppressive influence on early clearance of this pathogen from NALT. Keywords: disease state analysis

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Identification of Prx binding partners from Streptococcus pyogenes lysates

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