A simplified and easy-to-use HIP HOP assay provides insights into chalcone antifungal mechanisms of action

Thomas A.K. Prescott; Laura Anaissi-Afonso; Keith R. Fox; Anthony Maxwell; Barry Panaretou; Félix Machín

doi:10.1002/1873-3468.14483

Original language	English
Pages (from-to)	3087-3102
Number of pages	16
Journal	FEBS Letters
Volume	596
Issue number	23
DOIs	https://doi.org/10.1002/1873-3468.14483
Accepted/In press	2022
Published	Dec 2022
Additional links	Link to publication in Scopus

Bibliographical note

Funding Information: The authors thank Dominic Hoepfner of Novartis for advice on strain selection and Moses Langat for verification of compound identity using NMR. FM was supported by the Ministerio de Ciencia e Innovación (research grant BFU2017‐83954‐R); Agencia Canaria de Investigación, Innovación y Sociedad de la Información (research grant ProID2017010167, and predoctoral fellowship TESIS2017010039 to LA‐A); and Fundación Canaria Instituto de Investigación Sanitaria de Canarias (research grant PIFIIS19/04). BFU2017‐83954‐R, ProID2017010167 and TESIS2017010039 were co‐financed with the European Commission's ERDF Structural Funds. Work in AM's lab was supported by a Wellcome Trust Investigator Award (110072/Z/15/Z) and Biotechnology and Biosciences Research Council (BBSRC) Institute Strategic Programme Grant (BB/P012523/1). The authors thank Alison Howells (Inspiralis Ltd.) for carrying out the topoisomerase assays. Funding Information: The authors thank Dominic Hoepfner of Novartis for advice on strain selection and Moses Langat for verification of compound identity using NMR. FM was supported by the Ministerio de Ciencia e Innovación (research grant BFU2017-83954-R); Agencia Canaria de Investigación, Innovación y Sociedad de la Información (research grant ProID2017010167, and predoctoral fellowship TESIS2017010039 to LA-A); and Fundación Canaria Instituto de Investigación Sanitaria de Canarias (research grant PIFIIS19/04). BFU2017-83954-R, ProID2017010167 and TESIS2017010039 were co-financed with the European Commission's ERDF Structural Funds. Work in AM's lab was supported by a Wellcome Trust Investigator Award (110072/Z/15/Z) and Biotechnology and Biosciences Research Council (BBSRC) Institute Strategic Programme Grant (BB/P012523/1). The authors thank Alison Howells (Inspiralis Ltd.) for carrying out the topoisomerase assays. Publisher Copyright: © 2022 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.

Abstract

Elucidating the mechanism of action of an antifungal or cytotoxic compound is a time-consuming process. Yeast chemogenomic profiling provides a compelling solution to the problem but is experimentally complex. Here, we demonstrate the use of a highly simplified yeast chemical genetic assay comprising just 89 yeast deletion strains, each diagnostic for a specific mechanism of action. We use the assay to investigate the mechanism of action of two antifungal chalcone compounds, trans-chalcone and 4′-hydroxychalcone, and narrow down the mechanism to transcriptional stress. Crucially, the assay eliminates mechanisms of action such as topoisomerase I inhibition and membrane disruption that have been suggested for related chalcone compounds. We propose this simplified assay as a useful tool to rapidly identify common off-target mechanisms.

King's College London

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