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• W2766738034 endingPage "3966" @default.
• W2766738034 startingPage "3955" @default.
• W2766738034 abstract "Evolve and resequence experiments have provided us a tool to understand bacterial adaptation to antibiotics. In our previous work, we used short-term evolution to isolate mutants resistant to the ribosome targeting antibiotic kanamycin, and reported that Escherichia coli develops low cost resistance to kanamycin via different point mutations in the translation Elongation Factor-G (EF-G). Furthermore, we had shown that the resistance of EF-G mutants could be increased by second site mutations in the genes rpoD/cpxA/topA/cyaA Mutations in three of these genes had been discovered in earlier screens for aminoglycoside resistance. In this work, we expand our understanding of these second site mutations, the goal being to understand how these mutations affect the activities of the mutated gene products to confer resistance. We show that the mutation in cpxA most likely results in an active Cpx stress response. Further evolution of an EF-G mutant in a higher concentration of kanamycin than what was used in our previous experiments identified the cpxA locus as a primary target for a significant increase in resistance. The mutation in cyaA results in a loss of catalytic activity and probably results in resistance via altered CRP function. Despite a reduction in cAMP levels, the CyaAN600Y mutant has a transcriptome indicative of increased CRP activity, pointing to an unknown role for CyaA and / or cAMP in gene expression. From the transcriptomes of double and single mutants, we describe the epistasis between the mutation in EF-G and these second site mutations. We show that the large scale transcriptomic changes in the topoisomerase I (FusAA608E-TopAS180L) mutant likely result from increased negative supercoiling in the cell. Finally, genes with known roles in aminoglycoside resistance were present among the misregulated genes in the mutants." @default.
• W2766738034 created "2017-11-10" @default.
• W2766738034 creator A5046122126 @default.
• W2766738034 creator A5054645433 @default.
• W2766738034 creator A5079563104 @default.
• W2766738034 date "2017-12-01" @default.
• W2766738034 modified "2023-10-14" @default.
• W2766738034 title "Modulation of Global Transcriptional Regulatory Networks as a Strategy for Increasing Kanamycin Resistance of the Translational Elongation Factor-G Mutants in <i>Escherichia coli</i>" @default.
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• W2766738034 doi "https://doi.org/10.1534/g3.117.300284" @default.
• W2766738034 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5714492" @default.
• W2766738034 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/29046437" @default.

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