Functional Characterization Of Styr-3 Non-Protein-Coding Rna (npcRNA) In Salmonella enterica serovar Typhi
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Date
2014-12
Authors
Kumar, Thiviyaa M. Othaya
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Non-protein-coding RNAs (npcRNAs) are RNA molecules that do not
encode for proteins but are involved in modulating a myriad of cell regulatory
processes, including bacterial drug resistance mechanisms. In this study, a novel
npcRNA, StyR-3; discovered from the genome of S. Typhi was selected for further
characterization via bioinformatics and experimental-based approaches. This 144-nt
intergenic npcRNA is located in the ramRA regulon of S. Typhi Ty2. The
bioinformatics-based characterization showed that it is highly conserved among
Salmonella species; besides having a stable secondary structure as was predicted by
LocaRNA software. According to BPROM software, StyR-3 was predicted to
possess its own promoter. Furthermore, RNApredator; a target RNA search program
showed that StyR-3 could form a stable interaction with the ramR mRNA, which
further suggest its potential role in the regulation of MDR cascade. This was then
followed by cloning and overexpression of StyR-3 in an S. Typhi clinical strain
(ST001) in order to determine its effect upon the mRNA levels of ramA and ramR.
The Northern blot expression result suggest that the overexpression of StyR-3 does
not incur significant genotypic effect over the ramA and ramR mRNA levels.
Moreover, disk diffusion assay that was carried out to determine the phenotypic
effect of StyR-3 overexpression showed that both; the recombinant and the nonrecombinant
strains were susceptible to 7 out of 8 drugs tested. Collectively, these
results suggest that StyR-3 is most probably not involved in MDR regulation but it
could possibly exert other biological roles.
Description
Keywords
Characterization of STYR-3 non-protein-coding RNA , in Salmonella enterica serovar Typhi