Presentation is loading. Please wait.

Presentation is loading. Please wait.

Global changes in Staphylococcus aureus gene expression during human prosthetic joint infection AAU I n t r o d u c t.

Similar presentations


Presentation on theme: "Global changes in Staphylococcus aureus gene expression during human prosthetic joint infection AAU I n t r o d u c t."— Presentation transcript:

1 Global changes in Staphylococcus aureus gene expression during human prosthetic joint infection AAU I n t r o d u c t i o n M e t h o d s C o n c l u s i o n s R e s u l t s Identification Staphylococcus aureus is one of the leading causes of community- and hospital-acquired infections worldwide. It can cause acute infections and adapt to a biofilm mode of growth and thereby cause persistent and recurrent infections, particularly in device- related infections. Little is known about regulation of gene activity of S. aureus during actual human infection. Here we characterize the metabolome using NMR, and the transcriptome using RNA-seq, of S. aureus infected joint fluid derived from an acute human prosthetic joint infection, and compare them with the genome, transcriptome and metabolome of an isolate obtained from the same sample grown in vitro (LB medium). S. aureus sustained on a versatile human-cell-based diet consisting of amino acids, glycans and nucleosides in the hypoxic joint fluid during human prosthetic joint infection. Many, but not all, of the known virulence factor genes were upregulated in situ. Yijuan Xu 1, Raluca Georgiana Maltesen 1, Lone Heimann Larsen 1,2, Henrik Carl Schønheyder 2, Jeppe Lund Nielsen 1, Per Halkjær Nielsen 1, Trine Rolighed Thomsen 1,3, Kåre Lehmann Nielsen 1, The PRIS Study Group 3 1 Department of Biotechnology, Chemistry, and Environmental Engineering, Aalborg University, Denmark. 2 Department of Clinical Microbiology, Aalborg Hospital, Aalborg University Hospital, Denmark. 3 The Danish Technological Institute, Life Science Division, Denmark. To characterize the in situ virulence gene expression and metabolism of S. aureus using RNA-seq and NMR metabolite analysis. Gene expression Fig. 1 Overexpressed metabolic pathways in the infection. The pathway names are according to the MetaCyc database. Each pathway is assigned with a specific color and the upregulated enzymes in each pathway are indicated. A i m 1,3-Bisphospho-D-glycerate Dihydroxyacetone phosphateD-glyceraldehyde-3-phophate β-D-glucose-6-phosphate D-fructose-6-phosphate Fructose-1,6-bisphosphate 3-Phospho-D-glycerate 2-Phospho-D-glycerate Phosphoenolpyruvate gapA Phosphate NAD + H + NADH gpmA fbp Phosphate H2OH2O Pyruvate Acetyl-CoA formate pflB Coenzyme A Acetaldehyde Ethanol NADH NAD + H+H+ Coenzyme A NADH NAD + H+H+ aldA ADH adhP L-serine tdcB L-alanine H + ammonia NADH Ammonia 2 H + NAD + H 2 O ald Deoxyribose-1-phosphate Phosphate Adenine DeoD Phosphate Guanine DeoD 2-Deoxy-α-D-ribose 1-phosphate DeoB 2-Deoxy-D-ribose-5-phosphate DeoC Acetaldehyde AldA ADH Acetyl-CoA NAD + Coenzyme A NADH H + Phosphate Uracil DeoA Thymidine Phosphate Thymine DeoA Lactose 6-phosphate D-galactose 6-phosphate LacG Tagatose-6-phosphate NAD + H + NADH Coenzyme A LacA LacB H2OH2O b-D-glucose Tagatose-1,6-bisphosphate ADP H + LacC ATP LacD L-arginine H2OH2O Ammonia H + L-citrulline Phosphate H + Carbamoyl-phosphate CO 2 ADP H + Ammonia ATP L-ornithine arcA argI argF yqeA arcC L-proline Ammonium SAUREUSv1_20053 (S)-1-pyrroline-5-carboxylate L-glutamate SAUREUSv1_30035 rocA L-histidine Ammonia H + Urocanate hutH H + A reduced electron acceptor An oxidized electron acceptor 2 H 2 O NAD + H + NADH 4-Imidazolone-5-propionate hutU H 2 O H+H+ N-forminino-L-glutamate hutU H2OH+H2OH+ H 2 O Formate N-acetylneuraminate N-acetyl-β-D-mannosamine nanA L-arginine L-ornithine Lactose 6-phosphate LacE LacF arcD DeoxyguanosineDeoxyuridine Spontaneous (S)-2-acetolactate Spontaneous A reduced electron acceptor CO 2 An oxidized electron acceptor H + Diacetyl(S)-acetoin NAD + NADH H + butA PURINE/PYRIMIDINE DEOXYRIBONUCLEOSIDES DEGRADATION GLYCOLYSIS I LACTOSE AND GALACTOSE DEGRADATION I L-SERINE DEGRADATION ALANINE DEGRADATION IV PYRUVATE TO ETHANOL FERMENTATION I (S)-ACETOIN BIOSYNTHESIS N-ACETYLNEURAMINATE DEGRADATION ARGININE DEIMINASE PATHWAY ORNITHINE DEGRADATION I PROLINE DEGRADATION HISTIDINE DEGRADATION I Deoxyadenosine D) Nucleosides B) Carbohydrates A) Amino acids C) Fermentation products Fig. 2 Concentration of metabolites determined by NMR analysis. LB(OD 600 =0) (blue) and joint fluid (green) were analyzed in technical triplicates while LB (OD 600 =0.5) (red) was done in biological replicates. The detection limit of NMR is 2 μM. S. aureus monoinfection was determined by culture, 16S amplicon sequencing and FISH. 436 Protein-coding genes (17% of total) were differentially expressed (322 upregulated and 114 downregulated in situ). 131 Known or proposed virulence factors in the genome: 47 upregulated and 9 downregulated. Particularly γ-hemolysins, a few superantigen-like proteins, adhesins and immune evasion molecules as well as SaeRS and VraSR two-component systems were overexpressed in situ. A c k n o w l e d g e m e n t This paper was prepared within the framework of the ‘Prosthetic-Related Infection and Pain’ (PRIS) - Innovation project. Thehttp://www.joint-prosthesis-infection-pain.dk study was supported by a grant for the PRIS Innovations Consortium from The Danish Council for Technology and Innovation (no. 09–052174). Culture FISH 16S amplicon sequencing Synovial fluid Isolate (OD 600 ~0.5) RNA-seq Bioinformatics (EdgeR) Genome sequencing and annotation Synovial fluid Isolate (OD 600 ~0.5) NMR measurement Metabolite analysis


Download ppt "Global changes in Staphylococcus aureus gene expression during human prosthetic joint infection AAU I n t r o d u c t."

Similar presentations


Ads by Google