结核分枝杆菌(Mtb),引起结核病的病原体,表现出不同的遗传谱系,与毒力有已知联系。虽然已经探索了现代和古代Mtb谱系之间的基因组和转录组差异,小的非编码RNA(sRNA)在翻译后基因调控中的作用在很大程度上仍然未知.在这项研究中,Mtb谱系1(L1)Sabahan菌株(n=3)进行了sRNA测序,揭示351个sRNAs,包括23个已知的sRNAs和328个使用ANNOgesic鉴定的新的sRNAs。基于300的最佳平均截断值选择了13种sRNA,RT-qPCR揭示了MtbL1和其他谱系(L2和L4,n=3)之间sRNA1(p=0.0132)和sRNA29(p=0.0012)的显着表达差异(p>0.05)。使用RACE(cDNA末端的快速扩增)进一步表征,随后用TargetRNA3进行靶标预测,揭示了sRNA1(55个碱基对)靶向Rv0506、Rv0697和Rv3590c,和sRNA29(86个碱基对)靶向Rv33859c,Rv3345c,Rv0755c,Rv0107c,Rv1817,Rv2950c,Rv1181,Rv3610c,和Rv3296。Mycobrowser的功能表征揭示了这些参与调节中间代谢和呼吸的目标,细胞壁和细胞过程,脂质代谢,信息途径,和PE/PPE。总之,两个新的sRNAs,sRNA1和sRNA29在L1和其他谱系之间表现出差异表达,在基本的Mtb功能中具有预测的作用。这些发现提供了对Mtb调控机制的见解,有望在未来发展改进的结核病治疗策略。
Mycobacterium tuberculosis (Mtb), the tuberculosis-causing agent, exhibits diverse genetic lineages, with known links to virulence. While genomic and transcriptomic variations between modern and ancient Mtb lineages have been explored, the role of small non-coding RNA (sRNA) in post-translational gene regulation remains largely uncharted. In this study, Mtb Lineage 1 (L1) Sabahan strains (n = 3) underwent sRNA sequencing, revealing 351 sRNAs, including 23 known sRNAs and 328 novel ones identified using ANNOgesic. Thirteen sRNAs were selected based on the best average cut-off value of 300, with RT-qPCR revealing significant expression differences for sRNA 1 (p = 0.0132) and sRNA 29 (p = 0.0012) between Mtb L1 and other lineages (L2 and L4, n = 3) (p > 0.05). Further characterization using RACE (rapid amplification of cDNA ends), followed by target prediction with TargetRNA3 unveils that sRNA 1 (55 base pairs) targets Rv0506, Rv0697, and Rv3590c, and sRNA 29 (86 base pairs) targets Rv33859c, Rv3345c, Rv0755c, Rv0107c, Rv1817, Rv2950c, Rv1181, Rv3610c, and Rv3296. Functional characterization with Mycobrowser reveals these targets involved in regulating intermediary metabolism and respiration, cell wall and cell processes, lipid metabolism, information pathways, and PE/PPE. In summary, two novel sRNAs, sRNA 1 and sRNA 29, exhibited differential expression between L1 and other lineages, with predicted roles in essential Mtb functions. These findings offer insights into Mtb regulatory mechanisms, holding promise for the development of improved tuberculosis treatment strategies in the future.