Analysis and Exploitation of Oxygen-Dependent Modification to Ribosomes

Lead Research Organisation: University of Oxford
Department Name: Oxford Chemistry

Abstract

Ribosomes are intracellular machines that make proteins, but they also contain proteins. Recent BBSRC funded work led to the discovery that ribosomal proteins undergo a modification, termed hydroxylation, dependent on atmospheric oxygen at sites of importance for protein biosynthesis. One of the largest classes of antibiotics, e.g. erythromycin, targets ribosomes. Point mutations to ribosomes confer resistance to such antibiotics. Hence it should be possible to develop compounds that selectively target unmodified over modified ribosomes. Tumour cells are often hypoxic so should contain ribosomes that are ales hydroxylated than normal cells. We propose to use Follow on Funding to test synthetically modified antibiotics (guided by ribosome structures) to identify compounds that selectively inhibit unmodified over modified (or vice versa) ribosomes. The results will be used to support patent applications and, if appropriate, to form a spin-out company.

Publications


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Katz MJ (2014) Sudestada1, a Drosophila ribosomal prolyl-hydroxylase required for mRNA translation, cell homeostasis, and organ growth. in Proceedings of the National Academy of Sciences of the United States of America
Loenarz C (2014) Hydroxylation of the eukaryotic ribosomal decoding center affects translational accuracy. in Proceedings of the National Academy of Sciences of the United States of America
Markolovic S (2015) Protein Hydroxylation Catalyzed by 2-Oxoglutarate-dependent Oxygenases. in The Journal of biological chemistry
 
Description The key technical objectives of the work have all been achieved, i.e. we optimised mass-spectrometric methods for the analysis of ribosomal proteins, developed new methods for the synthesis of modified antibiotics targeting ribosomes (new collaboration with Professor Darren Dixon, Department of Chemistry, Oxford, which has resulted in a publication on synthesis), and carried out cell based studies with a range of antibiotics to test the hypothesis that ribosomal proteins differentially modified by oxygenases would show different sensitivities to antibiotics binding close to the location of the modification (hydroxylation). These results are presently being written up for publication and, maybe, a patent application. Furthermore, the methods that we have developed have proven to be useful in our basic science work on post-translational modifications catalysed by oxygenases.
Exploitation Route Most of the work has focused on obtaining reliable data on prokaryotic cells, but we have also initiated work on human cells, the preliminary results of which will form the basis of a future large-scale application in the cancer field.
Sectors Chemicals,Healthcare,Pharmaceuticals and Medical Biotechnology
 
Description The work has impacted on antimicrobial resistance research (by defining a new type of oxygenase catalysed resistance mechanism). The methods developed will also be useful in research targeting prokaryotic oxygenases (for antibacterial resistance) and human oxygenases for diseases ranging from cancer. Specifically, they enable the quantitative mass spectrometric based analyses of the activities of oxygenases to be measured in a cellular context. The methods are useful both in terms of developing inhibition of oxygenases acting on ribosomes, and of measuring 'off target' inhibition of such oxygenases by inhibitors targeting other oxygenases, e.g. the hypoxia inducible factor prolyl hydroxylases for the treatment of anaemia, or the JmjC histone demethylases for the treatment of cancer.
Sector Chemicals,Healthcare,Pharmaceuticals and Medical Biotechnology
Impact Types Societal
 
Description ELRIG Meeting, Telford 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Talk at The European Laboratory Research & Innovation Group Meeting in Telford on "Targeting Oxygenases for the Regulation of Protein Biosynthesis"
Year(s) Of Engagement Activity 2015
 
Description GRC Enzyme, coenzymes and Metabolic Pathways, Waterville, USA 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation Gordon Research Conference, Waterville, USA on "The Enzymology of Oxygen Sensing in Humans and Other Organisms"
Year(s) Of Engagement Activity 2014
 
Description Gordon Research Conference, Ventura, California, USA 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation at Gordon Research Conference, Ventura, California, USA on "Roles of Oxygenases in Hypoxia Sensing"
Year(s) Of Engagement Activity 2015
 
Description Informa Life Sciences' Enzymes Event, London 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Talk at Informa Life Sciences' Enzymes Event, London on "Protein Hydroxylation by Oxygenases"
Year(s) Of Engagement Activity 2015
 
Description Lilly Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation at Lilly's Chemical Biology meets Drug Discover conference on "Chemical Biology of Oxygen Sensing"
Year(s) Of Engagement Activity 2014
 
Description Oxford Transcription Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Talk at Oxford Transcription Meeting on "Studies on the regulation of transcription by oxygen"
Year(s) Of Engagement Activity 2015
 
Description Proteomics Meeting COST, Oxford 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Talk at Proteomics Meeting COST, Oxford on "Proteomics for Functional Assignment of Oxygenases (is Hard Work)"
Year(s) Of Engagement Activity 2015
 
Description Seminar at Plant Sciences 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Seminar on 'Oxygenases as Hypoxia Sensors in Humans, Plants, and Other Organisms'
Year(s) Of Engagement Activity 2015
 
Description Society of Chemical Industry's London Region Committee, New York University London 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Industry/Business
Results and Impact Talk at Society of Chemical Industry's London Region Committee, New York University London on "The chemistry of oxygen sensing in humans and bacteria (from antibiotics to epigenetics and back again)"
Year(s) Of Engagement Activity 2014