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Streptococcus pyogenes M49 591

ABSTRACT: Serotype M49 strain associated with skin infection

PROVIDER: PRJNA13286 | ENA |

REPOSITORIES: ENA

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			Action	DRS
		Other
	SRR14857119_1.fastq.gz	Fastqsanger.gz
	SRR14857120_1.fastq.gz	Fastqsanger.gz
	SRR14857120_2.fastq.gz	Fastqsanger.gz

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Streptococcus pyogenes M49 591

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Identification of small non coding RNAs in S. pyognes M49 using intergenic tiling arrays

Project description:Small non-coding RNAs (sRNAs) have attracted attention as a new class of gene regulators in eukaryotes and in bacteria. In this study, we identified novel sRNAs of the human pathogen Streptococcus pyogenes M49 (GAS M49). A temporal expression profile of potential sRNAs was determined for (GAS M49). Cells were grown in chemical defined medium (CDM), Todd-Hewitt broth (Invitrogen) supplemented with 0.5% yeast extract (THY) or Brain-Heart-Infusion (BHI) complex medium using tiling arrays representing the intergenic regions of the GAS M49 genome. We identified 55 putative sRNAs in GAS M49 that were expressed during growth. Of those, 35 sRNAs were novel, whereas 20 sRNAs were known. Already described sRNAs included molecules belonging to one of the common non-coding RNA families covered by the rfam collection and streptococcal sRNAs that have been detected in previous studies. Comparison to a recently published bioinformatics screen showed a low overlap between putative sRNA genes. This is in accordance with the results from other studies, which underlines the fact that a comprehensive analysis of sRNAs expressed by a given organism requires the complementary use of different methods and the investigation of several environmental conditions. Despite a high conservation of sRNA genes within streptococci, the expression of sRNAs is rather strain specific.

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Galleria mellonella immune response to infection with Streptococcus pyogenes

Project description:The larvae of Galleria mellonella are increasingly popular as infection model organism to study virulence mechanisms and pathogenicity of bacteria and fungi. In this study, the Streptococcus pyogenes-specific immune response of the larvae was investigated using a proteomics approach. Larvae of the greater wax moth G. mellonella were obtained from Bugs-International GmbH (Irsingen, Germany). The larvae were stored and fed in a wheat bran mixture at 37°C. Larvae with a weight of 0.15 – 0.2 g were used in the experiments. S. pyogenes M49 strain 591 was grown overnight in THY, washed twice in a 0.9% NaCl solution, and suspended in 0.9% NaCl to a final concentration of 2.4 – 4 x 108 CFU/mL. Larvae were infected with 2.4 – 4 x 106 CFU/larva in 10 µL 0.9% NaCl. In each experiment, 45 larvae were infected with S. pyogenes and 30 larvae were injected with 0.9 % NaCl (mock control). Hemolymph was extracted after 4 h, 24 h, and 72 h from 15 infected larvae and 10 non-infected larvae per time point. The hemolymph was separated by centrifugation into cell-free hemolymph and hemocytes. Each experiment was performed in three biological replicates. The protein abundance was regulated in the course of the infection. In the early infection phase, four hours post infection (p.i.), proteins responsible for recognition of foreign surfaces were upregulated, e.g. peptidoglycan recognition proteins. After 24 h, sensory proteins, receptors involved in recognition and opsonisation, and antimicrobial peptides were detected with increased amounts compared to the control group. 72 h p.i., proteins involved in development and metabolism were present at reduced levels. Overall, the immune response of larvae showed relative specificity to S. pyogenes infection, high similarity to the mammalian innate immune response, and a greater complexity than appreciated today.

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