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Biology 240 Course-Based Undergraduate Research Experience

Rifampicin Resistance Mutation and Fitness Cost


Jeffrey Taylor-Kantz, DiShawnn Newell, Camille Georges

BACKGROUND METHODS COST OF RESISTANCE


Rifampicin is an antibiotic that • Selected for resistance • Natural selection will act upon E. coli that have Rif-
resistance, making them more likely to survive and
binds to the beta subunit of RNA • Growth of bacteria via transfers reproduce.
polymerase and inhibits • MIC assessment (level of resistance) • However, this higher fitness is due to the
transcription. However, if the shape • Competitive Fitness Assay (growth rate) conformational change of the E. coli's RNA
polymerase, specifically at the active binding site
of the RNA polymerase changes via • PCR DNA Sequencing of Rifampicin. Although this shape change will lead
a mutation, rifampicin becomes to less binding of rifampicin to the RNA, making
the bacteria to be more resistant, it will also lead
less likely to bind and the bacteria to the RNA Polymerase being less likely to bind to
can now survive in this adverse the DNA strand and proceed with transcription.
• This lack of transcription will lead to less
environment. The RpoB gene is translation of proteins. Eventually, the bacteria will
responsible for the expression of perish due to inability to perform life functions.
the E. Coli RNA polymerase, and a
mutation within this gene could CONCLUSIONS & NEXT STEPS
possibly increase the bacteria's • Resistance-causing mutation must have
occurred somewhere else on the chromosome
level of resistance to rifampicin.
• Reverted to sensitivity (progenitor was more
Rifampacin binding resistant than the descendent)
UW CURE OVERVIEW pocket • This supports the hypothesis that resistance
has a fitness tradeoff cost
• No collateral effects observed with
RESULTS streptomycin
• Next Step: sequence other portions of the
No mutations were observed in RpoB gene in search of mutations that could
either have caused resistance and research collateral
effects among other drugs
Cluster I nor Cluster II
ACKNOWLEDGEMENTS
MIC FITNESS DATA • Thank you to Elizabeth Glenski, Matt Seivers, and Grace Dy for designing
the template poster format.
• Special Thanks to Jeff Smith, lab TA super star
Progenitor: 3240 µg/mL Progenitor: 1.1739 • Thank you to our dedicated team of Peer Facilitators – Bao Nguyen, Deja
Machen, Komalpreet Brar, Maia Nguyen, Jess Xiao, Rachel Hu, and Cailin
Descendent: 13 µg/mL Descendent: 2.2650 Winston
Figure 1: CURE Experimental Design Layout. • We would also like to thank the Katie Dickenson and the CURE prep team.