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A chemical potentiator of copper-accumulation used to investigate the iron-regulons of Saccharomyces cerevisiae

Lookup NU author(s): Dr Andrew Foster, Dr Sam Dainty, Carl Patterson, Dr Julian Rutherford, Professor Nigel Robinson

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

The extreme resistance of Saccharomyces cerevisiae to copper is overcome by 2-(6-benzyl-2-pyridyl)quinazoline (BPQ), providing a chemical-biology tool which has been exploited in two lines of discovery. First, BPQ is shown to form a red (BPQ)(2)Cu(I) complex and promote Ctr1-independent copper-accumulation in whole cells and in mitochondria isolated from treated cells. Multiple phenotypes, including loss of aconitase activity, are consistent with copper-BPQ mediated damage to mitochondrial iron-sulphur clusters. Thus, a biochemical basis of copper-toxicity in S. cerevisiae is analogous to other organisms. Second, iron regulons controlled by Aft1/2, Cth2 and Yap5 that respond to mitochondrial iron-sulphur cluster status are modulated by copper-BPQ causing iron hyper-accumulation via upregulated iron-import. Comparison of copper-BPQ treated, untreated and copper-only treated wild-type and fra2 Delta by RNA-seq has uncovered a new candidate Aft1 target-gene (LSO1) and paralogous non-target (LSO2), plus nine putative Cth2 target-transcripts. Two lines of evidence confirm that Fra2 dominates basal repression of the Aft1/2 regulons in iron-replete cultures. Fra2-independent control of these regulons is also observed but CTH2 itself appears to be atypically Fra2-dependent. However, control of Cth2-target transcripts which is independent of CTH2 transcript abundance or of Fra2, is also quantified. Use of copper-BPQ supports a substantial contribution of metabolite repression to iron-regulation.


Publication metadata

Author(s): Foster AW, Dainty SJ, Patterson CJ, Pohl E, Blackburn H, Wilson C, Hess CR, Rutherford JC, Quaranta L, Corran A, Robinson NJ

Publication type: Article

Publication status: Published

Journal: Molecular Microbiology

Year: 2014

Volume: 93

Issue: 2

Pages: 317-330

Print publication date: 09/07/2014

Online publication date: 15/06/2014

Acceptance date: 29/05/2014

ISSN (print): 0950-382X

ISSN (electronic): 1365-2958

Publisher: Wiley-Blackwell

URL: http://dx.doi.org/10.1111/mmi.12661

DOI: 10.1111/mmi.12661


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