A chemical potentiator of copper‐accumulation used to investigate the iron‐regulons of S accharomyces cerevisiae
Summary The extreme resistance of S accharomyces 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 Ctr 1‐independen...
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Published in | Molecular microbiology Vol. 93; no. 2; pp. 317 - 330 |
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Main Authors | , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
01.07.2014
|
Online Access | Get full text |
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Summary: | Summary
The extreme resistance of
S
accharomyces 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
Ctr
1‐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
Aft
1/2,
Cth
2 and
Yap
5 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
Δ by
RNA
‐seq has uncovered a new candidate
Aft
1 target‐gene (
LSO1
) and paralogous non‐target (
LSO2
), plus nine putative
Cth
2 target‐transcripts. Two lines of evidence confirm that
Fra
2 dominates basal repression of the
Aft
1/2 regulons in iron‐replete cultures.
Fra
2‐independent control of these regulons is also observed but
CTH2
itself appears to be atypically
Fra
2‐dependent. However, control of
Cth
2‐target transcripts which is independent of
CTH2
transcript abundance or of
Fra
2, is also quantified. Use of copper‐
BPQ
supports a substantial contribution of metabolite repression to iron‐regulation. |
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ISSN: | 0950-382X 1365-2958 |
DOI: | 10.1111/mmi.12661 |