Poster abstracts
Poster number 117 submitted by Yow Yong Tan
Investigating the importance of the bulge region for S. enterica htrA RNA thermometer behavior using SHAPE assays.
Yow Yong Tan (Department of Chemistry and Biochemistry, Denison University), Rachel M. Mitton-Fry (Department of Chemistry and Biochemistry, Denison University)
Abstract:
RNA thermometers (RNATs) are structural elements that can be found in the 5′- untranslated region of bacterial mRNAs that encode for heat shock proteins or virulence factors. They regulate the expression of these genes by changing conformation in response to temperature. Such regulation enables immediate response to environmental changes and allows pathogenic bacteria to detect the successful invasion of a warm-blooded host. In Salmonella enterica, an RNA thermometer in the htrA gene regulates the expression of HtrA, an enzyme critical for Salmonella virulence. Our previous work showed that a single-nucleotide bulge plays a significant role in tuning the htrA thermometer melting behavior. To better our understanding of the importance of the bulge region, we studied the melting behavior of a mutant htrA thermometer that switched the position of the base pairs flanking the bulge. We performed Selective 2′-Hydroxyl Acylation analyzed by Primer Extension (SHAPE) assays on the mutant and wild-type at different temperatures representing the standard growth conditions, warm-blooded body temperatures, and heat-shock conditions. Our results reveal that this mutation seems to have a subtle effect on the melting behavior of the thermometer.
Keywords: SHAPE, RNAT, htrA