Transcriptomics,Genomics

Dataset Information

21

Aerobic and Anaerobic Growth of Streptococcus pneumoniae D39


ABSTRACT: The catalase-negative, facultative anaerobe Streptococcus pneumoniae D39 is naturally resistant to hydrogen peroxide (H2O2) produced endogenously by pyruvate oxidase (SpxB). Here, we investigate the adaptive response to endogenously produced H2O2. We show that lactate oxidase, which converts lactate to pyruvate, positively impacts pyruvate flux through SpxB and that ∆lctO mutants produce significantly lower H2O2. In addition, both the SpxB and pyruvate dehydrogenase complex (PDHC) pathways contribute to acetyl-CoA production during aerobic growth, and the pyruvate format lyase (PFL) pathway is the major acetyl-CoA pathway during anaerobic growth. Microarray analysis of the D39 strain cultured under aerobic vs. strict anaerobic conditions show up-regulation of spxB, a rhodanese-like protein (spd0091), tpxD, sodA, piuB, piuD and an Fe-S protein biogenesis operon under H2O2-producing conditions. Proteome profiling of H2O2-induced sulfenylation reveals that sulfenylation levels correlate with cellular H2O2 production, with endogenous sulfenylation of ≈50 proteins. Deletion of tpxD increases cellular sulfenylation 5-fold and has an inhibitory effect on ATP generation. Two major targets of protein sulfenylation are glyceraldehyde-3-phosphate dehydrogenase (GapA) and SpxB itself, but also include pyruvate kinase, LctO, AdhE and acetate kinase (AckA). Sulfenylation of GapA is inhibitory, while the effect on SpxB activity is negligible, consistent with this cell-abundant protein functioning as a “sink” for endogenous H2O2. Strikingly, four enzymes of capsular polysaccharide biosynthesis are sulfenylated, as are enzymes associated nucleotide biosynthesis via ribulose-5-phosphate. We propose that LctO/SpxB-generated H2O2 functions as a signaling molecule to down-regulate capsule production and drive altered flux through sugar utilization pathways. Overall design: Three independent hybridizations using independent RNA preparations from the strain IU1690 (replicate 1 and 2) and strain IU1781 (replicate 3) were performed. Strain IU1781 is isogenic with IU1690 except for the rpsL1 mutation. Reference samples were grown aerobically while experimental samples were grown anaerobically. Dye swap was performed with the reference strain labeled with Cy3 in two hybridizations and Cy5 in one hybridizations.

INSTRUMENT(S): MWG Streptococcus pneumoniae Array

SUBMITTER: Ho-Ching Tiffany Tsui   

PROVIDER: GSE19791 | GEO | 2016-12-08

SECONDARY ACCESSION(S): PRJNA121813

REPOSITORIES: GEO

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Publications

Biological and Chemical Adaptation to Endogenous Hydrogen Peroxide Production in Streptococcus pneumoniae D39.

Lisher John P JP   Tsui Ho-Ching Tiffany HT   Ramos-Montañez Smirla S   Hentchel Kristy L KL   Martin Julia E JE   Trinidad Jonathan C JC   Winkler Malcolm E ME   Giedroc David P DP  

mSphere 20170104 1


The catalase-negative, facultative anaerobe Streptococcus pneumoniae D39 is naturally resistant to hydrogen peroxide (H2O2) produced endogenously by pyruvate oxidase (SpxB). Here, we investigate the adaptive response to endogenously produced H2O2. We show that lactate oxidase, which converts lactate to pyruvate, positively impacts pyruvate flux through SpxB and that ΔlctO mutants produce significantly lower H2O2. In addition, both the SpxB pathway and a candidate pyruvate dehydrogenase complex (  ...[more]

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