Title of Abstract

Elucidating the Effect of the Antibiotic Tetracycline on the Regulatory Function of the Guanidine-Sensing ykkCD Riboswitch

Poster Number

21

Faculty Mentor

Timea Fernandez, Ph.D.; fernandezt@winthrop.edu

College

College of Arts and Sciences

Faculty Mentor

Timea Fernandez, Ph.D.

Abstract

The primary objective for one of several project avenues spearheaded by the Fernandez lab is to develop an assay - ideally one more feasible for small-to-medium sized and/or undergraduate-only scientific research institutions - for evaluating the concomitant binding of tetracycline (tet antibiotic) and guanidine (guanidinium cation at physiologic pH) to the guanidine-sensing bacterial riboswitch ykkCD. Thus more generally a guanidine-I riboswitch, this noncoding regulatory RNA segment has been shown to also recognize the translation-inhibiting antibiotic via a high-affinity aptamer domain. Resultant conformational change in the regulator prompts translation-wise upregulation of genes downstream from it, among which - of those known - most encode membrane-bound transporters (e.g., SMR efflux pumps) responsible for expelling such toxins. This phenomenon establishes a major underpinning of the seemingly ever-evolving issue of antibiotic resistance in bacteria, further evincing that, aside from having to "rely on" incomparably slower multigenerational evolution of purely novel tactics, modern bacteria quickly adapt and employ their diverse detoxification machinery to render various traditional therapeutic agents largely ineffective. A binding assay is planned to decipher how proximal or distal one ligand-riboswitch binding region is to/from the other and to investigate relevant thermodynamics & kinetics, through which it should be distinguishable whether or not the influence of tet-ykkCD association on guanidine recognition is statistically significant. Soil bacterium Bacillus subtilis is the RNA source herein, whereas intestinal bacterium Escherichia coli is the replication vehicle/host (vector). Past prescribed assays call for cumbersome procedures, including radioisotope-labeled RNA utilization and tedious gel sequencing. The assay under development therefore seeks to employ more universally accessible methodology, ideally both isothermal titration calorimetry (ITC) and surface plasmon resonance spectroscopy (SPR), for tet-influenced guanidine-regulator binding affinity and efficiency determination.

Additional Fields About Your Abstract

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Course Assignment

CHEM 551 and CHEM 552 - Fernandez

Other Presentations/Performances

Fall 2020 SURE Showcase at Winthrop University, Rock Hill, SC 29733, October 2020

Grant Support

NIH-NCBI, SC EPSCoR, and INBRE

Type of Presentation

Poster presentation

Start Date

16-4-2021 11:30 AM

SullivanT10_551_Literature Review.docx (102 kB)
Fernandez Research - Literature Review (Summer 2020)

SullivanT10_Poster_SURE2020_Fernandez.pptx (605 kB)
Fernandez Research - Poster (year-round)

SullivanT10_551_updated_Presentation_shortened.pptx (6592 kB)
Fernandez Research - CHEM 551/552 Presentation draft (year-round)

group presentation copy1.pptx (39663 kB)
Gelabert Research - Slides: SOURCE/SURE 2019-2020 (Please allow submission for personal records purposes)

updatedMarch2020_Sullivan_Thomas_2020SOURCEAbstract.docx (23 kB)
Gelabert Research - Abstract: SOURCE/SURE 2019-2020 (Please allow submission for personal records purposes)

long version --- Thomas Sullivan poster - Research on Hydrothermal-based Discovery of New Compounds in K-La-Zr-O Quaternary System.pptx (9603 kB)
Gelabert Research - Poster: SOURCE/SURE 2019-2020 (Please allow submission for personal records purposes)

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Apr 16th, 11:30 AM

Elucidating the Effect of the Antibiotic Tetracycline on the Regulatory Function of the Guanidine-Sensing ykkCD Riboswitch

The primary objective for one of several project avenues spearheaded by the Fernandez lab is to develop an assay - ideally one more feasible for small-to-medium sized and/or undergraduate-only scientific research institutions - for evaluating the concomitant binding of tetracycline (tet antibiotic) and guanidine (guanidinium cation at physiologic pH) to the guanidine-sensing bacterial riboswitch ykkCD. Thus more generally a guanidine-I riboswitch, this noncoding regulatory RNA segment has been shown to also recognize the translation-inhibiting antibiotic via a high-affinity aptamer domain. Resultant conformational change in the regulator prompts translation-wise upregulation of genes downstream from it, among which - of those known - most encode membrane-bound transporters (e.g., SMR efflux pumps) responsible for expelling such toxins. This phenomenon establishes a major underpinning of the seemingly ever-evolving issue of antibiotic resistance in bacteria, further evincing that, aside from having to "rely on" incomparably slower multigenerational evolution of purely novel tactics, modern bacteria quickly adapt and employ their diverse detoxification machinery to render various traditional therapeutic agents largely ineffective. A binding assay is planned to decipher how proximal or distal one ligand-riboswitch binding region is to/from the other and to investigate relevant thermodynamics & kinetics, through which it should be distinguishable whether or not the influence of tet-ykkCD association on guanidine recognition is statistically significant. Soil bacterium Bacillus subtilis is the RNA source herein, whereas intestinal bacterium Escherichia coli is the replication vehicle/host (vector). Past prescribed assays call for cumbersome procedures, including radioisotope-labeled RNA utilization and tedious gel sequencing. The assay under development therefore seeks to employ more universally accessible methodology, ideally both isothermal titration calorimetry (ITC) and surface plasmon resonance spectroscopy (SPR), for tet-influenced guanidine-regulator binding affinity and efficiency determination.