The transcriptome responses to hydrogen peroxide, H 2 O 2 , of the facultatively phototrophic bacterium Rhodobacter sphaeroides grown under semiaerobic conditions were investigated. At 7 min after the addition of 1 mM H 2 O 2 , the expression of approximately 9% of all genes (total, 394) was changed reliably by at least twofold. At 30 min, the number of genes (total, 88) and the magnitude of expression changes were much lower, indicating rapid recovery from stress. Two types of responses were observed: (i) an H 2 O 2 stress response per se and (ii) a shift to high-oxygen metabolism. The former response involved the upregulation of genes for H 2 O 2 detoxification, protein folding and proteolysis, DNA damage repair, iron transport and storage, iron-sulfur cluster repair, and the downregulation of genes for protein translation, motility, and cell wall and lipopolysaccharide synthesis. The shift to high-oxygen metabolism was evident from the differential regulation of genes for aerobic electron transport chain components and the downregulation of tetrapyrrole biosynthesis and photosystem genes. The abundance of photosynthetic complexes was decreased upon prolonged exposure of R. sphaeroides to H 2 O 2 , thus confirming the physiological significance of the transcriptome data. The regulatory pathways mediating the shift to high-oxygen metabolism were investigated. They involved the anaerobic activator FnrL and the antirepressor-repressor AppA-PpsR system. The transcription of FnrL-dependent genes was down at 7 min, apparently due to the transient inactivation by H 2 O 2 of the iron-sulfur cluster of FnrL. The transcription of the AppA-PpsR-dependent genes was down at 30 min, apparently due to the significant decrease in appA mRNA.In natural environments, microorganisms have to cope with oxidative stress caused by reactive oxygen species (ROS). In aerobically growing bacteria, the autooxidation of the respiratory chain components is believed to be one of the main sources of endogenous ROS (16,20,30); however, additional sources exist (48). ROS are also produced by exposure of aerobically grown cells to metals, redox-active chemicals, or radiation. To cope with damage inflicted by ROS, bacteria induce oxidative stress defense systems. These include superoxide dismutases, catalases, and peroxidases directly involved in ROS detoxification and repair mechanisms. The ROS-oxidized cysteine and methionine residues of proteins are repaired (reduced) by the thioredoxin, glutathion/glutaredoxin, and methionine sulfoxide reductase systems. DNA is protected from damage by specific ROS-induced DNA-binding proteins, while damaged DNA is repaired via the SOS response (1, 8, 52).Rhodobacter sphaeroides is a facultatively phototrophic bacterium belonging to the alpha division of the proteobacteria that can use various pathways for energy generation. At high O 2 tension, it uses aerobic respiration and lacks photosynthetic complexes. When O 2 tension drops, the expression of the photosystem (PS) genes increases, resulting in the produ...