Robo-Therm, a pipeline to RNA Thermometer discovery and validation.
Davis M ShartsMaria T AlmanzaAndrea Victoria BanksAlyssa M CastellanosCatherine G O HernandezMonica L LopezDaniela RodriguezAlina Y TongMaximilian R SegebergLuiz F M PassalacquaMichael M AbdelsayedPublished in: RNA (New York, N.Y.) (2024)
RNA thermometers are highly structured noncoding RNAs located in the 5' untranslated regions (UTR) of genes that regulate expression by undergoing conformational changes in response to temperature. The discovery of RNA thermometers through bioinformatics is difficult because there is little sequence conservation among their structural elements. Thus, the abundance of these thermo-sensitive regulatory structures remains unclear. Herein, to advance the discovery and validation of RNA thermometers, we developed Robo-Therm, a pipeline that combines an adaptive and user-friendly in silico motif search with a well-established reporter system. Through our application of Robo-Therm, we discovered two novel RNA thermometers in bacterial and bacteriophage genomes found in the human gut. One of these thermometers is present in 5'-UTR of a gene that codes for σ70 RNA polymerase subunit in the bacteria Mediterraneibacter gnavus and Bacteroides pectinophilus, and in the bacteriophage Caudoviricetes, which infects Bacteroides pectinophilus. The other thermometer is in the 5'-UTR of a tetracycline resistance gene (tetR) in the intestinal bacteria Escherichia coli and Shigella flexneri. Our Robo-Therm pipeline can be applied to discover multiple RNA thermometers across various genomes.
Keyphrases
- escherichia coli
- genome wide
- high throughput
- endothelial cells
- gene expression
- copy number
- genome wide identification
- crispr cas
- multidrug resistant
- pseudomonas aeruginosa
- staphylococcus aureus
- molecular dynamics
- mass spectrometry
- cystic fibrosis
- long non coding rna
- microbial community
- induced pluripotent stem cells
- antibiotic resistance genes
- clinical evaluation