Loss of RNase J leads to multi-drug tolerance and accumulation of highly structured mRNA fragments in Mycobacterium tuberculosis.
Loss of RNase J leads to multi-drug tolerance and accumulation of highly structured mRNA fragments in Mycobacterium tuberculosis.
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核糖核酸酶J的缺失导致结核分枝杆菌对多药耐药和高结构的mRNA片段积聚。
DOI：
10.1371/journal.ppat.1010705
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发表时间：
2022-07
期刊：
影响因子：
6.7
通讯作者：
中科院分区：
文献类型：
--
作者：
尽管存在赋予结核分枝杆菌（Mtb）高水平耐药性的良好表征的典型突变，但有证据表明耐药机制比简单获得此类突变更复杂。最近的研究表明，结核分枝杆菌可以获得非典型的耐药相关突变，在某些药物的存在下赋予生存优势，可能作为获得高水平耐药的垫脚石。编码RNase J的Rv 2752 c/rnj在耐药临床Mtb分离株中不成比例地突变。在这里，我们表明，rnj的删除赋予增加耐受致死浓度的几种药物。RNAseq揭示了RNase J影响富含PE/PPE基因和稳定RNA的基因子集的表达，并且是适当的23 S rRNA成熟的关键。基因表达差异暗示两种sRNA和ppe 50-ppe 51是药物耐受性表型的重要贡献者。此外，我们发现在没有RNase J的情况下，许多短RNA片段积累，因为它们以较慢的速率降解。我们发现，积累的转录片段是RNase J的目标，其特征在于强大的二级结构和高G+C含量，这表明RNase J在降解高度结构化的RNA中具有限速作用。综上所述，我们的结果表明，RNase J间接影响药物耐受性，以及揭示RNase J在分枝杆菌RNA代谢中的内源性作用。结核分枝杆菌是导致结核病（TB）的细菌，每年造成100多万人死亡。几种抗生素对结核病有效。然而，M。结核病经常发生突变，导致抗生素耐药性，使治疗变得困难，有时甚至不可能。为了制定更好的战略来对抗耐药结核病，我们需要了解耐药性是如何获得的。研究揭示了“踏脚石突变”的存在，可能导致细菌具有低水平的抗生素耐药性，使一些细菌能够在治疗中存活并获得导致高水平抗生素耐药性的额外突变。RNase J是一种参与RNA加工和降解的细菌酶，先前发现其突变与M.结核我们假设这些可能是垫脚石突变，因此研究了RNase J与耐药性之间的关系。我们发现RNase J的缺失导致更多的M。结核病细胞在抗生素治疗中存活。我们确定这种存活率的增加是由于在没有RNase J的情况下发生的基因表达的变化。这项工作很重要，因为它描述了一种细菌可以用来逃避抗生素治疗的新机制。
Despite the existence of well-characterized, canonical mutations that confer high-level drug resistance to Mycobacterium tuberculosis (Mtb), there is evidence that drug resistance mechanisms are more complex than simple acquisition of such mutations. Recent studies have shown that Mtb can acquire non-canonical resistance-associated mutations that confer survival advantages in the presence of certain drugs, likely acting as stepping-stones for acquisition of high-level resistance. Rv2752c/rnj, encoding RNase J, is disproportionately mutated in drug-resistant clinical Mtb isolates. Here we show that deletion of rnj confers increased tolerance to lethal concentrations of several drugs. RNAseq revealed that RNase J affects expression of a subset of genes enriched for PE/PPE genes and stable RNAs and is key for proper 23S rRNA maturation. Gene expression differences implicated two sRNAs and ppe50-ppe51 as important contributors to the drug tolerance phenotype. In addition, we found that in the absence of RNase J, many short RNA fragments accumulate because they are degraded at slower rates. We show that the accumulated transcript fragments are targets of RNase J and are characterized by strong secondary structure and high G+C content, indicating that RNase J has a rate-limiting role in degradation of highly structured RNAs. Taken together, our results demonstrate that RNase J indirectly affects drug tolerance, as well as reveal the endogenous roles of RNase J in mycobacterial RNA metabolism. Mycobacterium tuberculosis is the bacterium that causes tuberculosis (TB), which kills over a million people each year. Several antibiotics are effective against TB. However, M. tuberculosis frequently acquires mutations that cause antibiotic resistance, making treatment difficult and sometimes impossible. To develop better strategies to combat antibiotic-resistant TB, we need to understand how resistance is acquired. Studies have revealed the presence of “stepping-stone mutations” that may cause bacteria to have low levels of antibiotic resistance, allowing some bacteria to survive treatment and acquire additional mutations that cause high levels of antibiotic resistance. Mutations in RNase J, a bacterial enzyme involved in the processing and degradation of RNA, were previously found to be associated with antibiotic resistance in M. tuberculosis. We hypothesized that these could be stepping-stone mutations and therefore investigated the relationship between RNase J and drug resistance. We found that deletion of RNase J causes more M. tuberculosis cells to survive antibiotic treatment. We determined that this increased survival is due to changes in gene expression that occur in the absence of RNase J. This work is important because it describes a new mechanism that bacteria can use to escape antibiotic treatment.
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影响因子：
14.9
作者：
Hnilicová J;Jirát Matějčková J;Šiková M;Pospíšil J;Halada P;Pánek J;Krásný L
通讯作者：
Krásný L
影响因子：
6.4
作者：
DeJesus MA;Gerrick ER;Xu W;Park SW;Long JE;Boutte CC;Rubin EJ;Schnappinger D;Ehrt S;Fortune SM;Sassetti CM;Ioerger TR
通讯作者：
Ioerger TR
影响因子：
3.2
作者：
Daniel, J;Deb, C;Kolattukudy, PE
通讯作者：
Kolattukudy, PE
影响因子：
5.7
作者：
Dorleans, Audrey;de la Sierra-Gallay, Ines Li;Condon, Ciaran
通讯作者：
Condon, Ciaran
影响因子：
2.8
作者：
Gengenbacher, Martin;Rao, Srinivasa P. S.;Dick, Thomas
通讯作者：
Dick, Thomas