利福平对耻垢分枝杆菌基因的调控  

Regulation of Rifampicin on Genes of Mycobacterium Smegmatis

作　　者：刘通 杨丽[1] 陈赢 唐静 林源[1] 马国荣 杨延辉[1] LIU Tong;YANG Li;CHEN Ying;TANG Jing;LIN Yuan;MA Guorong;YANG Yanhui(School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院,银川750004

出　　处：《宁夏医科大学学报》2021年第5期461-468,共8页Journal of Ningxia Medical University

基　　金：国家自然科学基金(81560604);中国科学院“西部之光”人才培养引进计划的资助(XAB2019AW11);宁夏医科大学校级课题(XY201527)。

摘　　要：目的探讨利福平(RIF)对耻垢分枝杆菌(MS MC2155)转录组表达的影响。方法以RIF的1/2最小抑菌浓度(MIC)处理的MS MC2155菌株为实验组,用相同稀释倍数的DMSO处理MS MC2155菌株作为对照组,于摇床内37℃培养72 h后,用Trizol试剂提取各组MS MC2155的总RNA,通过Illumina Hiseq 3000测序平台进行全转录组测序,采用Bowtie 2软件比对测序结果,用MS MC2155标准菌株的参考基因组对样本基因进行注释,通过倍数差异分析,筛选获得RIF作用前后的差异基因(different expression gene,DEG),最后用GO及KEGG数据库注释上述获得的DEGs。结果DEG分析显示,在RIF处理后发现了832个DEGs(P≤0.05,|log2FC|≥1),其中457个基因上调(54.93%),375个基因下调(45.07%)。GO富集分析结果显示,DEGs(nuoF、nuoB和sigM)主要涉及氧化还原酶活性、呼吸链复合物等细胞与分子方面的功能,以及外来生物刺激、共生体调节等生物学过程。KEGG富集分析显示,DEGs(cspA、sigM、nrdB、gmk、nuoF、nuoB和mysB)主要集中在万古霉素耐药、氧化磷酸化、核苷酸代谢和酪氨酸代谢等通路。STRING分析结果显示,共有371个基因编码的蛋白质存在相互作用,其中处于中心节点的蛋白的基因包括mysB、pheT和guaB。结论经转录组测序发现,RIF既能影响MS MC2155中sigM、mysB等RNA合成基因的表达;又能抑制其nrdB、gmk和guaB DNA合成基因的表达,进而阻断其DNA和蛋白的合成。Objective To explore the effect of rifampicin(RIF)on the expression level of the transcriptome of mycobacterium smegmatis(MS)MC2155.Methods The treatment group was MS MC2155 processed with 1/2 minimum inhibitory concentration(MIC)of RIF,and the control group was MS MC2155 processed with the same concentration of DMSO,both two were cultured for 72 h.The total RNA was extracted from the two groups by Trizol,and transcriptomes were sequenced by Illumina Hiseq platform.The results were blasted with the reference genome of MS MC2155 by Bowtie 2.The Difference Expressed Genes(DEGs)were compared between the two groups.The DEGs obtained was annotated with GO and KEGG databases.Results There were 832 DEGs appeared after RIF intervention(P≤0.05,|log2FC|≥1).In these DEGs,457(54.93%)DEGs were up-regulated and 375(45.07%)DEGs were down-regulated.GO pathway enrichment analyses showed that DEGs,nuoF,nuoB and sigM,were mainly involved in oxidoreductase activity,respiratory chain complex and other cellular and molecular functions,as well as foreign biological stimulation,symbiote regulation and other biological processes.By KEGG pathway enrichment analyses,the majority of DEGs,cspA,sigM,nrdB,nuoF,nuoB and mysB,were involved in vancomycin resistance,oxidative phosphorylation,nucleotide and tyrosine metabolism.STRING analysis uncovered that 371 proteins interacted with each other,and mysB,pheT,guaB were included in the central nodes.Conclusion By transcriptome sequence we discovered that RIF not only disturbs key genes of RNA synthesis pathway,such as sigM,mysB,but also directly inhibits the essential genes of DNA synthesis,such as nrdB,gmk and guaB.Finally,RIF blocked the synthesis of mycobacterial DNA and proteins.

关 键 词：利福平 耻垢分枝杆菌 转录组 差异基因 结核病 耐药性 

分 类 号：R37[医药卫生—病原生物学] R91[医药卫生—基础医学]