Abstract:This study aimed to screen the potential genes or pathways regulated by RseA in Mycobacterium smegmatis and to provide preliminary verification of the results. M. smegmatis strains overexpressing RseA or a plasmid control were constructed through molecular cloning technology. Transcriptome sequencing and bioinformatics methods were used to compare the differentially expressed genes (DEGs) between the groups; to assess the DEG functional enrichment through gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis; and to construct a protein-protein interaction (PPI) network. M. tuberculosis resuscitation factor E (RpfE) was used to promote the recovery of non-replicating retained bacteria, and the resuscitation index (RI) was measured to perform preliminary verification of the screened key targeted genes or pathways. A total of 2 403 DEGs in the RseA overexpression group relative to the control were identified, comprising 2 335 up-regulated genes and 68 down-regulated genes. GO functional enrichment analysis showed that the DEGs mainly participated in the following categories: regulation of nitrogen compound metabolic process, RNA metabolic process, transcription factor activity and sequence-specific DNA binding. KEGG pathway analysis revealed enrichment in the pathways of terpenoid backbone biosynthesis, fructose and mannose metabolism and homologous recombination. A screen of the top eight hub genes (rpsG, rplM, rpsO, rpsT, rpmH, rplS, rpsJ and rplT) from the MCODE module of the PPI network revealed that these genes are involved in ribosome pathways. This study revealed that the RseA can regulate multiple targeted genes, affect the ribosome structure and ribosome pathways, and enhance the sensitivity of M. smegmatis to anti-tuberculosis drugs. These findings lay a foundation for further mechanistic research.
熊雨菡, 李智颖, 卢楠, 陈俊伊, 唐佳玲, 徐蕾, 杨春, 何永林. RseA基因影响耻垢分枝杆菌药物敏感性的生物信息学分析与初步验证[J]. 中国人兽共患病学报, 2021, 37(2): 115-121.
XIONG Yu-han, LI Zhi-ying, LU Nan, CHEN Jun-yi, TANG Jia-ling, XU Lei, YANG Chun, HE Yong-lin. Bioinformatics analysis and preliminary verification of the antibiotic sensitivity of Mycobacterium smegmatis, as affected by the RseA gene. Chinese Journal of Zoonoses, 2021, 37(2): 115-121.
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