GSE115993
Next Generation Sequencing Analysis of S. aureus cultured in the presence/absence of V12CBD
We reported the application of NGS technology for high-throughput transcription profiling of S. aureus in the presence of V12CBD, the binding domain from bacteriophage lysin PlyV12. In summary, a total of 225 differentially expressed genes (DEGs) were identified (at least 2-fold change, with a p-value <0.05) after V12CBD treatment. Of those, transcripts of 111 genes were found upregulated and 114 downregulated in response to V12CBD. Predominantly genes involved in translation (76.9%), nucleotide metabolism (80%), cell division (100%), as well as replication and repair (82%), were repressed in S. aureus treated with V12CBD. The GO enrichment analysis showed that the DEGs were scattered in 34 GO terms, specifically, a total of 3 items related to cellular components, 25 items for biological processes, and 6 items for molecular functions. Functional classification of the DEGs using KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis, revealed that a large group of S. aureus genes expressed differentially in response to V12CBD belonged to the membrane transport pathway, the environmental information processing category. Several genes encoding important virulence factors, such as surface protein encoding gene sdrC, protein arginine kinase encoding gene mcsB, persistent infection associated genes (putP and oppF2), and the genes involved in capsular polysaccharide biosynthesis (tarI and SAOUHSC_00125), were downregulated in the presence of V12CBD. Finally, two two-component systems (TCSs), WalKR, SrrAB, were also found downregulated in the presence of V12CBD. The transcription levels of these genes were also confirmed by primer-specific qRT-PCR. This study showed that V12CBD could suppress multiple important virulence factors expression in S. aureus. Overall design: S. aureus T23 was cultured in the presence and absence of 25 µg/ml V12CBD for 4 h at 37°C, and the mRNA profiles were generated by deep sequencing using Illumina NextSeq.