Characterisation of a putative oxidoreductase in Escherichia coli: YdgJ

Antibiotic resistance is one of the main challenges faced by humanity in the 21st century. In 2016, the Bolt lab identified by a genetic screen an uncharacterised putative oxidoreductase called YdgJ that conferred resistance to the antibiotic nalidixic acid in Escherichia coli. This protein was isol...

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Main Author: Cusin, Lola M.L.
Format: Thesis (University of Nottingham only)
Language:English
Published: 2020
Subjects:
Online Access:https://eprints.nottingham.ac.uk/60610/
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author Cusin, Lola M.L.
author_facet Cusin, Lola M.L.
author_sort Cusin, Lola M.L.
building Nottingham Research Data Repository
collection Online Access
description Antibiotic resistance is one of the main challenges faced by humanity in the 21st century. In 2016, the Bolt lab identified by a genetic screen an uncharacterised putative oxidoreductase called YdgJ that conferred resistance to the antibiotic nalidixic acid in Escherichia coli. This protein was isolated and purified for in vitro study, and further genetic analysis of YdgJ was carried out in parallel. The genetic analysis revealed that the YdgJ gene did not convey nalidixic resistance, contrary to earlier experiments. Surprisingly, expression of YdgJ protein caused increased sensitivity to ROS. We predicted that YdgJ would bind to cofactor NAD(P), but this could not be confirmed experimentally. We gained evidence that the YdgJ protein formed a trimeric structure, in agreement with predictions from bioinformatics. Overall, the mechanism of action of YdgJ could not be uncovered, but several clues hinted towards a role in ROS detoxification or cell metabolism. Further experiments will be required to reach a full understanding of the YdgJ gene and its associated product.
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format Thesis (University of Nottingham only)
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spelling nottingham-606102025-02-28T14:55:02Z https://eprints.nottingham.ac.uk/60610/ Characterisation of a putative oxidoreductase in Escherichia coli: YdgJ Cusin, Lola M.L. Antibiotic resistance is one of the main challenges faced by humanity in the 21st century. In 2016, the Bolt lab identified by a genetic screen an uncharacterised putative oxidoreductase called YdgJ that conferred resistance to the antibiotic nalidixic acid in Escherichia coli. This protein was isolated and purified for in vitro study, and further genetic analysis of YdgJ was carried out in parallel. The genetic analysis revealed that the YdgJ gene did not convey nalidixic resistance, contrary to earlier experiments. Surprisingly, expression of YdgJ protein caused increased sensitivity to ROS. We predicted that YdgJ would bind to cofactor NAD(P), but this could not be confirmed experimentally. We gained evidence that the YdgJ protein formed a trimeric structure, in agreement with predictions from bioinformatics. Overall, the mechanism of action of YdgJ could not be uncovered, but several clues hinted towards a role in ROS detoxification or cell metabolism. Further experiments will be required to reach a full understanding of the YdgJ gene and its associated product. 2020-07-31 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/60610/1/To%20upload.pdf Cusin, Lola M.L. (2020) Characterisation of a putative oxidoreductase in Escherichia coli: YdgJ. MRes thesis, University of Nottingham. Antibiotic resistance Escherichia coli Molecular microbiology
spellingShingle Antibiotic resistance
Escherichia coli
Molecular microbiology
Cusin, Lola M.L.
Characterisation of a putative oxidoreductase in Escherichia coli: YdgJ
title Characterisation of a putative oxidoreductase in Escherichia coli: YdgJ
title_full Characterisation of a putative oxidoreductase in Escherichia coli: YdgJ
title_fullStr Characterisation of a putative oxidoreductase in Escherichia coli: YdgJ
title_full_unstemmed Characterisation of a putative oxidoreductase in Escherichia coli: YdgJ
title_short Characterisation of a putative oxidoreductase in Escherichia coli: YdgJ
title_sort characterisation of a putative oxidoreductase in escherichia coli: ydgj
topic Antibiotic resistance
Escherichia coli
Molecular microbiology
url https://eprints.nottingham.ac.uk/60610/