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Gyrase Mutation in Various Bacteria
Leading to Fluoroquinolone (“Cipro”)
Resistance
By: Nancy Halliday
Wesley Hanson
Brenda Breeding
Audience:
Undergraduate Science Students
Major Objectives
• Understand the relationships between
DNA, RNA, and proteins.
• Explore the effects of tertiary protein
structural changes on enzyme function.
• Learn to utilize tools available in
bioinformatics.
Brief Overview
:
Students are presented journal articles discussing
fluoroquinolone resistance in various bacteria as a
result of Gyrase A mutation.
Students seek to determine the caused of primary
structure changes that lead to resistance. (Amino
acid substitutions within the enzyme Gyrase A are explored.)
Students search www.ncbi.nlm.nih.gov for the
appropriate protein code (1AB4) of the enzyme
“gyrase”
Brief Overview(cont.):
Students then utilize “Protein Explorer” to:
1. Visually examine the 3D structure of Gyrase A
2. Manipulate the program to identify the amino
acid substitutions that are identified as the point
of mutation(s).
3. Manipulate the program showing the structural
change in the molecule once the mutation has
taken place.
DNA Gyrase A and B
Functional Dimer
Ser 83
Gyrase A Subunit
Ser 83
Asp 87
Gyrase A Subunit
Brief Overview(cont.):
Students utilize “Biology Workbench” to:
1. Compare wildtype vs. mutated amino acid
sequence.
2. Compare wildtype vs. mutated nucleic acid
sequence.
Sequences are available in the primary literature
Blank
• Insert slide of DNA/Amino Acid sequence
alignments
Project Materials:
•Article: Rupp, J. et. al., Serine to asparagine
substitution in the gyrase A gene leads to
quinolone resistance in moxifloxacin-exposed
Chlamydia pneumoniae.
•Website: www.ncbi.nlm.nih.gov
•Software: “Protein Explorerer”
•Software: “Biology Workbench”
Future Directions
This initial exercise is used to introduce the
problem space and tools available.
Students are then assigned a specific
antibiotic resistance model to explore
mutations and protein tertiary structure
changes that result from these mutations.
Students then present results in a
professional format.