Tan Xinq Yuan, Citartan Marimuthu, Chinni Suresh Venkata, Ahmed Siti Aminah, Tang Thean-Hock
Advanced Medical and Dental Institute (AMDI), Universiti Sains Malaysia, Bertam, 13200 Kepala Batas, Penang Malaysia.
Department of Biotechnology, Faculty of Applied Sciences, AIMST University, 08100 Bedong, Kedah Malaysia.
Indian J Microbiol. 2023 Mar;63(1):33-41. doi: 10.1007/s12088-022-01050-9. Epub 2022 Dec 20.
Regulatory small RNAs (sRNA) are RNA transcripts that are not translated into proteins but act as functional RNAs. Pathogenic Leptospira cause an epidemic spirochaetal zoonosis, Leptospirosis. It is speculated that Leptospiral sRNAs are involved in orchestrating their pathogenicity. In this study, biocomputational approach was adopted to identify Leptospiral sRNAs. In this study, two sRNA prediction programs, i.e., RNAz and nocoRNAc, were employed to screen the reference genome of serovar Lai. Out of 126 predicted sRNAs, there are 96 cis-antisense sRNAs, 28 trans-encoded sRNAs and 2 sRNAs that partially overlap with protein-coding genes in a sense orientation. To determine whether these candidates are expressed in the pathogen, they were compared with the coverage files generated from our RNA-seq datasets. It was found out that 7 predicted sRNAs are expressed in mid-log phase, stationary phase, serum stress, temperature stress and iron stress while 2 sRNAs are expressed in mid-log phase, stationary phase, serum stress, and temperature stress. Besides, their expressions were also confirmed experimentally via RT-PCR These experimentally validated candidates were also subjected to mRNA target prediction using TargetRNA2. Taken together, our study demonstrated that biocomputational strategy can serve as an alternative or as a complementary strategy to the laborious and expensive deep sequencing methods not only to uncover putative sRNAs but also to predict their targets in bacteria. In fact, this is the first study that integrates computational approach to predict putative sRNAs in serovar Lai.
The online version contains supplementary material available at 10.1007/s12088-022-01050-9.
调控性小RNA(sRNA)是不翻译成蛋白质而是作为功能性RNA的RNA转录本。致病性钩端螺旋体引发一种流行性螺旋体人畜共患病——钩端螺旋体病。据推测,钩端螺旋体sRNA参与调控其致病性。在本研究中,采用生物信息学方法鉴定钩端螺旋体sRNA。在本研究中,使用了两个sRNA预测程序,即RNAz和nocoRNAc,来筛选赖型血清群的参考基因组。在126个预测的sRNA中,有96个顺式反义sRNA、28个反式编码sRNA以及2个与蛋白质编码基因有义方向部分重叠的sRNA。为了确定这些候选sRNA是否在病原体中表达,将它们与我们RNA测序数据集生成的覆盖文件进行了比较。结果发现,7个预测的sRNA在对数中期、稳定期、血清应激、温度应激和铁应激条件下表达,而2个sRNA在对数中期、稳定期、血清应激和温度应激条件下表达。此外,还通过RT-PCR实验证实了它们的表达。这些经过实验验证的候选sRNA还使用TargetRNA2进行了mRNA靶标预测。综上所述,我们的研究表明,生物信息学策略不仅可以作为费力且昂贵的深度测序方法的替代或补充策略,用于发现假定的sRNA,还可以预测它们在细菌中的靶标。事实上,这是第一项整合计算方法来预测赖型血清群中假定sRNA的研究。
在线版本包含可在10.1007/s12088-022-01050-9获取的补充材料。