The gene fmt, encoding tRNA(fMet)-formyl transferase, is essential for normal growth of M. bovis, but not for viability.
The gene fmt, encoding tRNA(fMet)-formyl transferase, is essential for normal growth of M. bovis, but not for viability.
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DOI：
10.1038/s41598-017-15618-9
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发表时间：
2017-11-09
影响因子：
4.6
通讯作者：
Barkan D
中科院分区：
文献类型：
--
作者：
Vanunu M;Lang Z;Barkan D
结核分枝杆菌是一个主要的健康威胁，需要新的药物靶点。细菌中的蛋白质合成使用带有甲酰化甲硫氨酸残基的引发剂tRNAi。甲酰基转移酶基因tRNAfMet（fmt）的缺失可导致大肠杆菌严重的生长迟缓。coli和S. pneumoniae，而不是铜绿假单胞菌或S.金黄色。fmt在M.转座子文库分析，但这从未在任何分枝杆菌中进行过正式测试。我们在M中进行了fmt的靶向缺失。与Mtb-复合物（M.牛）。在这两种情况下，我们创建了一个部分二倍体菌株，通过两步等位基因交换或特异性噬菌体转导删除天然基因，然后去除互补基因以创建完全缺失突变体。In M.可以容易地产生完整缺失菌株。与此相反，在M. bovis-BCG，仅在孵育6周后才能产生完全缺失菌株，其世代时间比wt细菌长约2倍。我们的结果证实了该基因在致病性分支杆菌中的重要性，但由于缺失突变体是可行的，因此fMT作为药物靶点的有效性仍不清楚。我们的研究结果也反驳了以前的报道，fmt是必不可少的M。结核病综合征
Mycobacterium tuberculosis is a major health threat, necessitating novel drug targets. Protein synthesis in bacteria uses initiator tRNAi charged with formylated methionine residue. Deletion of the formylase gene, tRNAfMet-formyl transferase (fmt), causes severe growth-retardation in E. coli and in S. pneumoniae, but not in P. aeruginosa or S. aureus. fmt was predicted to be essential in M. tuberculosis by transposon library analysis, but this was never formally tested in any mycobacteria. We performed a targeted deletion of fmt in M. smegmatis as well as Mtb-complex (M. bovis). In both cases, we created a mero-diploid strain, deleted the native gene by two-step allelic exchange or specialized-phage transduction, and then removed the complementing gene to create full deletion mutants. In M. smegmatis a full deletion strain could be easily created. In contrast, in M. bovis-BCG, a full deletion strain could only be created after incubation of 6 weeks, with a generation time ~2 times longer than for wt bacteria. Our results confirm the importance of this gene in pathogenic mycobacteria, but as the deletion mutant is viable, validity of fmt as a drug target remains unclear. Our results also refute the previous reports that fmt is essential in M. tuberculosis-complex.
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影响因子：
3.2
作者：
MEINNEL, T;GUILLON, JM;BLANQUET, S
通讯作者：
BLANQUET, S
影响因子：
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Liu, Jun O.
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通讯作者：
Mukherjee, Kakoli
影响因子：
3.5
作者：
Barkan D;Stallings CL;Glickman MS
通讯作者：
Glickman MS