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The characterization of the potential iron-acquisition gene dUTPase in Bacillus anthracis

Gallegos, Kyle Ross
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2023-05-04
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Abstract
Bacillus anthracis is the causative agent of the fatal disease anthrax, and its virulence is of great interest due to its potential as a biological weapon. B. anthracis causes disease by both escaping immune defenses and acquiring nutrients. A necessary nutrient that pathogens must acquire from its host is iron. To discover novel genes essential for iron acquisition, we screened transposon mutants in iron-deficient media with hemoglobin as the sole source of iron. We further prioritized the mutants discovered in our in vitro screen by assessing for attenuated virulence using our in vivo G. mellonella infection model. We found one mutant that has a disruption in the first gene of a two-gene operon containing putative dUTPase and aminopeptidase genes known as 9F12 Tn. Neither of these genes have been previously linked to iron acquisition. To confirm the role of the dUTPase gene in the observed 9F12 Tn phenotype, we created an independent insertional mutant in the dUTPase gene (dUTPase IM). We found that both of our mutants, 9F12 Tn and dUTPase IM, could not use hemoglobin as a source of iron. We also found that G. mellonella injected with 9F12 Tn and dUTPase IM had higher survival rates than those injected with the parent strain. Our results indicate that the dUTPase gene is necessary for iron-acquisition and virulence in B. anthracis. This study furthers our understanding of iron acquisition in a bacterial pathogen and increases our knowledge of how B. anthracis causes disease.
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Subject
Biology
Bacillus anthracis
Virulence
Nutrient acquisition
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Biology