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bioRxiv · 10.1101/476564

Gentamicin B1 is not a minor gentamicin component with major nonsense mutation suppression activity

Baradaran-Heravi, A.·Williams, D. E.·Powell, D. A.·Balgi, A. D.·Andersen, R. J.·Roberge, M.
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Abstract

Nonsense mutations are single base substitutions that introduce a premature termination codon (PTC) preventing the formation of full-length protein. They are the causative mutations in about 10% of patients in a large number of rare genetic diseases. High concentrations of the antibiotic gentamicin can induce the incorporation of an amino acid at a PTC and formation of full-length protein, a process called PTC readthrough. Gentamicin is composed of several related aminoglycosides. We recently reported (doi/10.1073/pnas.1620982114) that the major gentamicin components that are responsible for its antibacterial activity showed weak to no PTC readthrough activity but that the minor component gentamicin B1 was a potent readthrough inducer. We have now determined that gentamicin B1 acquired from the sole supplier at the time the study was carried out was not gentamicin B1 but instead the closely related aminoglycoside G418. Gentamicin B1 recently became available from a second commercial source. Here, we provide nuclear magnetic resonance (NMR) assignment data for the two commercial compounds and verify only the second is indeed gentamicin B1. We show that gentamicin B1 lacks PTC readthrough activity in HDQ-P1 and DMS-114 cells homozygous for the TP53 R213X nonsense mutation, as well as in a cell-free translation assay.

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Baradaran-Heravi, A., Williams, D. E., Powell, D. A., Balgi, A. D., Andersen, R. J., Roberge, M.. 2018-11-23. Gentamicin B1 is not a minor gentamicin component with major nonsense mutation suppression activity. https://doi.org/10.1101/476564

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