Abstract:We investigate the different expression of toxin gene mazF3,6,9 and antitoxin gene mazE3,6,9 in the drug-resistance Mycobacterium tuberculosis,we used quantitative real-time polymerase chin reaction method to detect the expression level of toxin gene mazF3,6,9 and antitoxin gene mazE3,6,9 in M. tuberculosis (20 mono-resistance strains, 20 multidrug resistance strains and standard strain H37Rv).The differences of gene expression levels between groups were analyzed by one-way ANOVA. Contrasting with control group, toxin genes mazF6,9 were up-regulated expression levels both in mono-resistance (11.1519±22.31721;8.4306±17.97897) and multidrug resistance (4.6016±1.29018;6.9627±6.92948), had statistical significance (P<0.01), mazF3 expression levels had statistical significance neither in mono-resistance nor in multidrug resistance (P>0.05); antitoxin genes mazE3 was in down-expression level, and had statistical significance both in mono-resistance (0.3606±0.12527) and multidrug resistance (0.2016±0.16542) (P<0.01), mazE6 had no statistical significance (P>0.05)either in mono-resistance or multi drug resistance, mazE9 only in multidrug resistance(0.3989±0.37679) was in down-expression level, and has statistical significance (P<0.001). The toxin gene mazF6,9 and antitoxin gene mazE3,9 may participate in the drug-resistance formation of M. tuberculosis.
刘微, 赵继利, 屈艳琳, 谢婉莹, 袁俐. qRT-PCR检测结核分枝杆菌毒素-抗毒素系统mazE F的表达[J]. 中国人兽共患病学报, 2017, 33(2): 143-147.
LIU Wei, ZHAO Ji-li, QU Yan-lin, XIE Wan-ying, YUAN Li. Detection on expression levels of mazE F toxin-antitoxin system in Mycobacterium tuberculosis by qRT-PCR. Chinese Journal of Zoonoses, 2017, 33(2): 143-147.
[1] Chang JN,Ning DG. Identification of a pair of Toxin-antitoxin (TA) gene in the chromosome of cyanobacteria synechocystis sp. PCC6803. Microbiology China, 2009, 36(1):31-36 (in Chinese) 常家宁, 宁德刚. 蓝细菌PCC6803染色体上的一对毒素-抗毒素基因的鉴定[J]. 微生物学通报, 2009, 36(1):31-36. [2] Gerdes K, Christensen SK, Lubnerolesen A. Prokaryotic toxin-antitoxin stress response loci[J]. Nat Rev Microbiol, 2005, 3(5):371-382. [3] Yamaguchi Y, Inouye M. Regulation of growth and death in Escherichia coli by toxin-antitoxin systems[J]. Nat Rev Microbiol, 2011, 9(11):779-790. [4] Winther KS, Gerdes K. Enteric virulence associated protein VapC inhibits translation by cleavage of initiator tRNA[J]. Proc Nat Acad Sci, 2011, 108(18):7403-7407. [5] Mutschler H, Gebhardt M, Shoeman R L, et al. A novel mechanism of programmed cell death in bacteria by Toxin-antitoxin systems corrupts peptidoglycan synthesis[J]. PLoS Biol, 2011, 9(3):e1001033-e1001033. [6] Tan Q, Awano N, Inouye M. YeeV is an Escherichia coli Toxin that Inhibits Cell Division by Targeting the Cytoskeleton Proteins, FtsZ and MreB[J]. Mol Microbiol, 2011, 79(1):109-118. [7] Prysak M H, Mozdzierz C J, Cook A M, et al. Bacterial toxin YafQ is an endoribonuclease that associates with the ribosome and blocks translation elongation through sequence-specific and frame-dependent mRNA cleavage[J]. Mol Microbiol, 2009, 71(5):1071-1087. [8] Schuster C F, RalphBertram. Toxin-antitoxin systems are ubiquitous and versatile modulators of prokaryotic cell fate[J]. FEMS Microbiol Lett, 2013, 340(2):73-85. [9] Aizenman E, Engelbergkulka H, Glaser G. An Escherichia coli chromosomal "addiction module" regulated by guanosine [corrected] 3',5'-bispyrophosphate: a model for programmed bacterial cell death[J]. Proc Nat Acad Sci, 1996, 93(12):6059-6063. [10] Zhang Y, Zhang J, Hoeflich KP, et al. MazF cleaves cellular mRNAs specifically at ACA to block protein synthesis in Escherichia coli [J]. Mol Cell, 2003, 12(4):913-923. [11] Suzuki M, Mao L, Inouye M. Single protein production (SPP) system in Escherichia coli [J]. Nat Protoc, 2007, 2(7):1802-1810. [12] Suzuki M, Zhang J, Liu M, et al. Single protein production in living cells facilitated by an mRNA interferase[J]. Mol Cell, 2005, 18(2):253-261. [13] Ramage HR, Connolly LE, Cox JS. Comprehensive functional analysis of Mycobacterium tuberculosis toxin-antitoxin systems: implications for pathogenesis, stress responses, and evolution[J]. PLoS Genet, 2009, 5(12):1000767. [14] Singh R, Barry CE, Boshoff HI. The three RelE homologs of Mycobacterium tuberculosis have individual, drug-specific effects on bacterial antibiotic tolerance[J]. J Bacteriol, 2010, 192(5):1279-1291. [15] Keren I, Minami S, Rubin E, et al. Characterization and transcriptome analysis of Mycobacterium tuberculosis persisters.[J]. Mbio, 2011, 2(3):00100-00111. [16] Korch SB, Contreras HClark-Curtiss JE. Three Mycobacterium tuberculosis Rel toxin-antitoxin modules inhibit mycobacterial growth and are expressed in infected human macrophages[J]. J Bacteriol, 2009, 191(5):1618-1630. [17] Ramirez MV, Dawson CC, Crew R, et al. MazF6 toxin of Mycobacterium tuberculosis , demonstrates antitoxin specificity and is coupled to regulation of cell growth by a Soj-like protein[J]. BMC Microbiol, 2013, 13(1):986-991. [18] Han JS, Lee JJ, Anandan T, et al. Characterization of a chromosomal toxin-antitoxin, Rv1102c-Rv1103c system in Mycobacterium tuberculosis [J]. Biochem Biophys Res Commun, 2010, 400(3):293-298. [19] Xu Y, Zhang Z, Sun Z. Drug resistance to Mycobacterium tuberculosis : from the traditional Chinese view to modern systems biology[J]. Crit Rev Mycrobio, 2015,41(3):399-410. [20] Buts L, Lah J, Dao-Thi MH, et al. Toxin-antitoxin modules as bacterial metabolic stress managers[J]. Trends Biochem Sci, 2005, 30(12):672-679. [21] Rustad TR, Harrell MI, Liao R, et al. The enduring hypoxic response of Mycobacterium tuberculosis [J]. PloS One, 2008, 3(1) e1502. [22] Albrethsen J, Agner J, Piersma SR, et al. Proteomic profiling of Mycobacterium tuberculosis identifies nutrient-starvation-responsive toxin-antitoxin systems [J]. Mol Cell Proteomics, 2013, 12(5):1180-1191. [23] Zhang Y, Zhang J, Hara H, et al. Insights into the mRNA cleavage mechanism by MazF, an mRNA interferase[J]. J Biol Chem, 2005, 280(5):3143-3150. [24] Erental A, Sharon I, Engelbergkulka H. Two programmed cell death systems in, Escherichia coli : an apoptotic-Like death is inhibited by the mazEF-mediated death pathway[J]. PLoS Biol, 2012, 10(3):490-493.