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Cite this: Metallomics, 2011, 3, 1124–1129

www.rsc.org/metallomics MINIREVIEW
The oxidative stress of zinc deficiencyw
David J. Eide
Received 13th June 2011, Accepted 7th July 2011

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DOI: 10.1039/c1mt00064k

Zinc is an essential catalytic and structural cofactor for many enzymes and other proteins. While
Zn2+ is not redox active under physiological conditions, it has been known for many years that
zinc deficiency causes increased oxidative stress and, consequently, increased oxidative damage to
DNA, proteins, and lipids. These results have indicated that zinc plays an indirect antioxidant
role and that dietary inadequacy may contribute to human diseases such as cancer. Recent studies
are helping to identify the primary sources of oxidative stress in low zinc. In addition, through
studies of the model eukaryotic cell, Saccharomyces cerevisiae, we are now beginning to
understand the strategies cells use to limit this stress and reduce its damage.

Introduction Approximately 2 billion people on Earth consume insufficient

Zinc is required for the function of many proteins playing both
catalytic and structural roles. It has been estimated that zinc is
an essential cofactor for the function of as many as 10% of the
proteins encoded by the human genome.1 These include DNA
binding proteins such as zinc finger proteins and the p53
tumor suppressor protein and enzymes such as Cu/Zn super-
oxide dismutase. Zinc deficiency is a worldwide health problem.
Department of Nutritional Sciences, University of Wisconsin-Madison,
1415 Linden Drive, Room 340B, Madison, WI 53706-1571.
E-mail: eide@nutrisci.wisc.edu; Fax: 608-262-5860;
Tel: 608-263-1613
w This article is published as part of a themed issue on Metal Toxicity,
Guest Edited by Gregor Grass and Christopher Rensing.
David Eide received his BS in
Microbiology from the Uni-
versity of Minnesota in 1981
and his PhD in Molecular
Biology from the University
of Wisconsin-Madison in
1987. He was a postdoctoral
researcher at the Massachusetts
Institute of Technology from
1987–1990 and then at the
University of Utah in
1990–1991. He began his work
on zinc as an Assistant
Professor in the Department
David J. Eide of Biochemistry and Molecular
Biology at the University of
Minnesota-Duluth. He moved to the Nutritional Science
Department at the University of Missouri-Columbia in 1996
and to the Department of Nutritional Sciences at the University
of Wisconsin-Madison in 2004.
amounts of zinc.2 Within the US alone, it was estimated that
almost 10% of people are at risk of inadequate zinc intake.2
The great importance of zinc to cellular function indicates that
this widespread deficiency is likely to have major health
consequences. This prediction is supported by epidemiological
studies linking zinc deficiency to the increased risk of chronic
disease including cancer.3,4
Given the large number of different roles that zinc plays in
biology, it is difficult to assign deficits in specific zinc-dependent
functions as contributing to the etiology of human diseases.
However, one obvious possible link between zinc deficiency
and disease is through zinc’s action as an antioxidant. The
antioxidant function of zinc has been a mystery for many
years.5,6 Because zinc is not redox active under physiological
conditions, it is clear that the metal is not directly involved in
donating electrons to or accepting electrons from oxidant
molecules. Therefore zinc is not functioning as an antioxidant
per se. Therefore, the antioxidant properties of zinc are a result
of some indirect mechanism of action. This review explores the
current data regarding the potential sources of oxidative stress
in zinc deficiency and what cellular responses to zinc deficiency
occur to limit the oxidative stress and repair the damage that
results from it.
Oxidative stress in zinc deficiency and
its consequences
Several studies have demonstrated that zinc deficiency
increases the production of reactive oxygen species. Many of
these studies have focused on zinc deficiency in cultured cells.
For example, Oteiza et al.7 showed that growth of mouse 3T3
cells in low zinc results in increased reactive oxygen species
(ROS) as measured using the ROS-sensitive fluorescent probe
2,7-dichlorodihydrofluorescein (DCFH). Upon reaction with
oxidants such as O2 , H2O2, and hydroxyl radical,8 DCFH is

1124 Metallomics, 2011, 3, 1124–1129 This journal is c The Royal Society of Chemistry 2011
converted to the fluorescent 2,7-dichlorofluorescein (DCF)
form. Similarly, Ho and Ames9 found that ROS levels
increased in zinc deficient rat C6 glioma cells. The increased
ROS in rat glioma cells was accompanied by an increase in
oxidative DNA damage. The effects of this damage were likely
compounded by the loss of key DNA repair mechanisms. It
was also shown that zinc deficiency in lung fibroblasts,10 liver
stellate cells,11 IMR-32 neuroblastoma cells,12 prostate epithelial
cells,13 and neuronal PC12 cells14 all show increased levels of
ROS and oxidative damage when grown in low zinc.
Increased ROS and resulting damage has also been found in
experimental animals fed low zinc diets. Perhaps the first
report of these effects in animals was made by Sullivan
et al.15 who showed that zinc-deficient rats had increased lipid
peroxidation in lung microsomes. These results were con-
firmed and expanded to effects in other tissues in a series of
papers by Bray and colleagues in the mid-1980s16,17 and in
subsequent papers by several other groups.18–23 Extension of
these findings to human subjects has also been made; increased
oxidative stress and resulting damage has been documented in
humans with sub-optimal zinc intakes.24,25 These findings
support the proposal that zinc deficiency may be a contributing
factor to the progression of cancer.26,27
Finally, we have recently determined that the yeast
Saccharomyces cerevisiae also shows increased levels of ROS
when grown under zinc-limiting conditions.28–30 This result
demonstrates that the effect of low zinc on oxidative stress
occurs in both simple, single-cell eukaryotes and metazoans.
In addition, these results have allowed us to bring the power of
yeast genetics to bear on this fundamental issue of zinc
nutrition as described later in this review.
Possible sources of ROS in low zinc
A number of possible mechanisms have been proposed to explain
the increased oxidative stress in zinc-limited cells (Fig. 1). A few
of these mechanisms have been discussed in prior reviews6,26,31 so
those will only be briefly summarized here. First, increased
oxidative stress may arise from decreased activity of key anti-
oxidant enzymes. As one obvious example, Cu/Zn superoxide
dismutase (SOD) is a zinc-dependent enzyme whose activity may
be compromised under zinc deficiency. Second, Zn-metallothionein
has recently been shown to also have antioxidant activity
through the oxidation of metal-bound cysteine ligands in the
protein.32,33 Expression of many metallothioneins is induced
by zinc treatment possibly leading to an increased ability to
eliminate reactive oxygen species. Thus, in zinc-limited cells
where metallothionein expression is low, this antioxidant
function would be lost. As a third mechanism, it has been
proposed that zinc may compete with redox active metal ions
like Cu and Fe for binding to sites on proteins and other
cellular macromolecules.34,35 Such competition would inhibit
the site-specific production of oxygen radicals through metal-
catalyzed Fenton chemistry. Finally, zinc has been proposed
to bind to and protect free sulfhydryl groups in proteins. This
may be another way in which zinc normally plays an anti-
oxidant role and inhibits production of reactive oxygen species.
At this time, there is little evidence available to evaluate
these different hypotheses in vivo. One hypothesis that can be
This journal is c The Royal Society of Chemistry 2011
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Fig. 1 Summary of possible sources of reactive oxygen species in
zinc-limited cells. Processes on the left are induced by zinc and limit
reactive oxygen species (ROS) production or accumulation. Thus, low
zinc would decrease these activities and cause increased ROS.
Conversely, the processes on the right are inhibited by zinc and increase
ROS production. Low zinc would increase these processes and
increase ROS production.
eliminated as a possible cause in at least some cell types is loss
of SOD activity. In several studies in which low zinc causes
oxidative stress, it has been noted that SOD activity actually
increased in those cells, probably as a response to the increased
oxidative stress.9,10 One type of cell in which zinc limitation
did decrease SOD activity was yeast. Low zinc growth reduced
SOD activity by approximately 2-fold.30 However, when SOD
was overexpressed from a plasmid vector in zinc-limited cells
and activity was restored to zinc-replete levels, oxidative stress
still occurred. Thus, loss of SOD activity is not a primary
cause of oxidative stress in low zinc in yeast or mammalian
cells. This is not to say, however, that SOD activity is not
important for tolerating the oxidative stress of zinc deficiency.
To identify what antioxidant enzymes were important for low
zinc growth, yeast mutants lacking 28 different antioxidant
enzymes were analyzed for growth defects in low zinc.30 These
mutants disrupted peroxiredoxins, glutathione peroxidases,
catalases, and superoxide dismutases. Of these, only two
antioxidant enzymes were found to be critical for zinc-limited
growth, Tsa1, the major cytosolic peroxiredoxin, and Sod1,
the cytosolic Cu/Zn superoxide dismutase. Superoxide dismutase
degrades superoxide to hydrogen peroxide and the Tsa1
peroxiredoxin degrades hydrogen peroxide to water. The fact
that these two enzymes work sequentially and that both are
required for low zinc growth suggests that the initial form of
ROS generated in low zinc is superoxide.
Additional possible sources of oxidative stress in low zinc
include disruption of the mitochondrial electron transport
chain. Zinc is required for many functions in mitochondria
including matrix-localized metalloproteases that proteolytically
Metallomics, 2011, 3, 1124–1129 1125
cleave newly arrived proteins during their mitochondrial
maturation. Mitochondrial enzymes such as the yeast Adh3
alchohol dehydogenase and Leu9 a-isopropylmalate synthase
(for leucine synthesis) also require zinc as a cofactor. In
addition, cytochrome c oxidase of the electron transport chain
requires zinc bound to its Cox4 subunit for function. Mutation
of the zinc site in Cox4 disrupts the structural stability of the
complex and breaks the electron transport chain.36 Such breaks
result in increased generation of ROS from the release of
electrons to oxygen largely by coenzyme Q. It was recently
shown that the mitochondrial matrix contains a labile cationic
pool of low molecular weight zinc, primarily in the form of a
complex with some unknown small molecule ligand, and that
this pool is required for respiratory function.37 In addition, zinc
deficiency was found to decrease expression of components of
the electron transport chain in human lung fibroblasts suggesting
another mechanism through which this pathway can be
disrupted.10 Thus, zinc deficiency could disrupt the function
or expression of electron transport chain components and
increase oxidative stress in that way.
Another candidate source of ROS in low zinc is the pathway
of disulfide bond formation that occurs in the secretory path-
way. Recent studies have shown that the ER is a potent
generator of ROS.38,39 Newly synthesized proteins with
reduced thiols react with a protein disulfide isomerase (Pdi1) to
generate disulfide bonds. Two electrons are transferred to Pdi1
during this reaction that are then passed to the Ero1 FAD-
dependent oxidase and then to molecular oxygen to generate
H2O2 and/or O2 .40,41 Proteins with correct disulfide bonds
are stable due to their proper folding. However, incorrect
disulfide bonds are sensitive to reduction by GSH (generating
GSSG) regenerating the reduced thiol state.40,42 Protein mis-
folding in the ER leads to more incorrect disulfide bonds and
cycling via the GSH-dependent pathway.43 Thus, the oxidative
protein folding system is a major generator of ROS and
GSSG under ER stress conditions.42 Several observations
suggest that the ER is a source of ROS in low Zn. First and
foremost is the observation that loss of ER zinc transporter
function in yeast increases the sensitivity of cells to exogenous
oxidants.44 We also found that Zn deficiency increases GSSG
levels28 and note that oxidative protein folding is the primary
source of GSSG in cells.42 Third, Zn deficiency activates the
unfolded protein response (UPR) in both yeast and mamma-
lian cells, indicating protein misfolding is occurring in the
ER.45,46 As noted above, expression of misfolded proteins in
the ER causes oxidative stress.43
While NADPH is well recognized as an important anti-
oxidant for its role in supplying electrons to glutathione reductase
and thioredoxin reductase, it is also clear that NADPH can also
play a pro-oxidant role at least in regard to zinc deficiency.
NADPH oxidase activity has recently been implicated as the
source of ROS in zinc-limited neuronal cells. NADPH oxidases
generate O2 from O2 for the purpose of microbial killing in
phagocytic cells and also to provide moderate levels of ROS
for its role in cellular signaling.47 It was shown previously that
activation of the N-methyl-D-aspartate (NMDA) receptor
caused superoxide production by activation of the NADPH
oxidase.48 Because zinc is a known inhibitor of the NMDA
receptor,49 Aimo et al. addressed whether zinc limitation of
1126 Metallomics, 2011, 3, 1124–1129
neuronal cells increased ROS production through loss of this
inhibitory effect.14 When activated, the NMDA receptor
triggers downstream responses by mediating calcium influx.
These authors showed that zinc-limiting differentiated PC-12
neuronal cells in culture resulted in a rapid increase in DCF
fluorescence. During that treatment, the p67phox subunit of the
NADPH oxidase complex was recruited to the membrane
fraction indicating activation of oxidase activity. These effects
were associated with an increase in cytosolic calcium that was
blocked by the NMDA receptor antagonist MK-801. Calcium
can activate NADPH oxidase activity through the stimulation
of protein kinase C. Both MK-801 and the protein kinase C
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inhibitor Ro-320432 also blocked NADPH oxidase activation
and the increased ROS in low zinc. Finally, direct inhibition of
NADPH oxidase with apocyanin blocked ROS production.
Thus, in neuronal cells, hyperactivation of the NMDA receptor
appears to be the initial cause of increased oxidative stress in
low zinc. It remains to be seen if types of cells that lack the
NMDA receptor have NADPH oxidase activity activated by
other mechanisms in low zinc. In this regard, it is intriguing to
note that the findings of Kojima-Yuasa et al.11 support the
possibility that increased NADPH oxidase activity may
potentially be the source of ROS in zinc-deficient hepatic stellate
cells. These authors found that treatment of zinc-deficient stellate
cells with an oxidase inhibitor decreased ROS production.
The caveat with these experiments is that the inhibitor used,
diphenyliodonium chloride, is not specific to NADPH oxidases.
Lastly, a pro-oxidant role of NADPH was also suggested by
Bray and colleagues.16,17 These authors found that rat lung
microsomes isolated from zinc deficient rats generated more
free radicals in the presence of NADPH than did those
isolated from zinc-replete animals. The NADPH-dependence
and sensitivity to inhibition by carbon monoxide suggested
that the source of this oxidative stress was cytochrome P450.
More recently, it was found that the level of P450 CYP4F2
isozyme was elevated in liver from zinc deficient rats which
may explain, at least in part, this elevated ROS generation.21
Cellular responses to zinc deficiency to alleviate
oxidative stress
The primary mechanism cells use to prevent the oxidative
stress of zinc deficiency is to maintain zinc homeostasis. Cells
of all organisms have mechanisms that provide for a constant
cytosolic and organellar zinc status while extracellular or
dietary levels fluctuate.50–53 In general, these mechanisms
include the control of zinc uptake, zinc efflux from the cell,
sequestration within organelles, or zinc binding by proteins
such as metallothionein. One key player in zinc homeostasis
with particular relevance to oxidative stress and zinc deficiency
is the Zap1 transcription factor of yeast. Zap1 is a zinc-
responsive transcriptional activator that induces transcription
of its target genes in zinc-limited cells and whose activity is
shut off in zinc-replete cells.54 This regulation occurs primarily
by zinc binding directly to the activation domains of this
transcription factor which folds those domains into conforma-
tions that are unable to recruit co-activator proteins that
facilitate transcription initiation.
This journal is c The Royal Society of Chemistry 2011

Fig. 2 Mechanism regulating sulfate assimilation in response to
organic sulfur compounds and zinc. Repression of sulfate assimilation
in low zinc (–Zn) conserves NADPH for its antioxidant roles. See text
for further details. Names in all capital letters and italicized, e.g.
MET30, denote genes while names not italicized in which only the first
letter is capitalized, e.g. Zap1, denote proteins. Ub, ubiquitin; Met,
methionine; SAM, S-adenosylmethionine; Cys, cysteine; GSH,
glutathione.
We and others have sought to identify the various targets of
Zap1 to understand the strategies cells use to thrive under
zinc deficiency. Through microarray analysis and other
approaches, it has been estimated that Zap1 induces the
expression of B80 genes in low zinc.55–59 One of these target
genes, TSA1 encoding the major cytosolic peroxiredoxin in the
cell, is induced by Zap1. Peroxiredoxins are antioxidant
enzymes that degrade H2O2 to H2O through the reducing
equivalents provided by thioredoxin and NADPH-dependent
thioredoxin reductase. The TSA1 gene is expressed in zinc-
replete cell to high levels such that those cells contain
B400 000 molecules of the Tsa1 protein per cell. The TSA1
gene was induced 2–3-fold in zinc-limited wild type cells and
this induction was blocked by deletion of the Zap1 gene28
Protein levels increased to the same extent. In addition, TSA1
expression was induced 2–3-fold in zinc-replete cells expressing
a constitutively active allele of Zap1 and the TSA1 promoter
contains a functional Zap1 binding site that is required for
zinc-responsive regulation. Deletion of the TSA1 gene resulted
in a strain that is severely defective for zinc-limited growth,
emphasizing the importance of this antioxidant to growth
under these conditions. Zinc-limited wild type yeast show a
marked increase in DCF fluorescence that is greatly increased
in a tsa1 null mutant. In addition, the GSSG : GSH ratio
increased in zinc-limited cells and this too was greatly exacerbated
by loss of Tsa1 function. Regulation by Zap1 was clearly
important for this resistance to oxidative stress; when the
promoter of TSA1 was mutated such that the Zap1 binding
site was mutated to prevent induction in low zinc (but without
affecting basal expression), these promoter mutant cells also
showed increased ROS production and a perturbed intra-
cellular redox environment. Thus, the doubling of Tsa1
protein levels conferred by Zap1 induction is clearly important
for low zinc growth. It is also notable that the CTT1 gene,
which encodes the cytosolic isoform of catalase, also appears
to be directly regulated by Zap1.57 UTH1, a gene of unknown
function that has also been implicated in oxidative stress
resistance, is a Zap1 target as well.60 Thus, through Zap1,
yeast cells respond to zinc deficiency by inducing expression of
antioxidant enzymes that protect the cell from the oxidative
stress generated under those conditions. We consider this to be
a ‘‘proactive’’ strategy of oxidative stress resistance rather
This journal is c The Royal Society of Chemistry 2011
than a ‘‘reactive’’ mechanism because these antioxidant genes
are induced in response to the cause of the increased ROS, low
zinc, rather than in response to the ROS itself.
Another aspect of Zap1 regulation relevant to oxidative
stress in low zinc is the control of sulfate assimilation. The
assimilation of sulfate occurs through a multi-step pathway
resulting in the generation of methionine and cysteine.61,62
This pathway has a high demand for NADPH requiring 6
moles of NADPH for every one mole of methionine or
cysteine produced. Enzymes on this pathway include those
encoded by the MET3, MET14, and MET16 genes. In our
recent transcriptomics analysis of zinc-responsive gene expres-
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sion, we identified 36 genes that were repressed in zinc-limited
cells in a Zap1-dependent manner.29 These apparently Zap1-
repressed genes included MET3, MET14, and MET16. This
observation raised the question of how a transcriptional
activator protein that induces expression could repress expres-
sion of these genes. The answer lies in the regulatory mechan-
ism that controls these genes in response to sulfur amino acid
levels. When these amino acids are low, MET3, MET14, and
MET16 are all induced in expression by the Met4 transcription
factor (Fig. 2). Met4 activity is regulated at the level of protein
stability. When cells are replete with organic sulfur compounds
(methionine, cysteine, S-adenosylmethionine, glutathione),
Met4 is ubiquitinated and degraded by the proteasome.62
Ubiquitination and targeting for degradation is mediated by
the SCFMet30 ubiquitin ligase. SCFMet30 is a multisubunit
ubiquitin ligase that relies on the Met30 subunit for target
protein specificity. The connection between Zap1 and MET
gene expression is the Met30 subunit. We found that the
MET30 gene is a Zap1 target whose expression is upregulated
in low zinc.29 Increased levels of Met30 increase the activity of
SCFMet30 and cause the degradation of Met4, loss of MET
gene expression, and decreased sulfate assimilation. Consistent
with this model, we found that zinc-limited cells had markedly
reduced levels of free methionine and cysteine. When the site
of ubiquitination in the Met4 protein was mutated such that
this protein could no longer be down-regulated in low zinc, we
found that these cells had decreased NADPH, elevated
GSSG : GSH ratio, increased ROS production, and increased
sensitivity to external oxidants such as H2O2. These results
indicate that repression of sulfate assimilation occurs in zinc-
limited cells to conserve NADPH for its antioxidant functions.
Summary and future directions
It has been known for many years that zinc-deficient mamma-
lian cells in culture and whole animals on low zinc diets
produce increased levels of ROS. There is also ample evidence
that zinc limitation in human subjects results in oxidative
stress. This stress can lead to damage of proteins, lipids, and
DNA. DNA damage can lead to mutation and this may
explain the epidemiological link between zinc deficiency and
cancer and other chronic diseases. The ultimate source of the
oxidative stress of zinc deficiency remains a mystery. The
recent finding of elevated NADPH oxidase activity in zinc-
limited neuronal cells suggests one possible mechanism of
ROS production. It will be interesting to determine if this
mechanism also applies to cells of other types. Recent studies
Metallomics, 2011, 3, 1124–1129 1127
of yeast have established that even single-cell eukaryotes
experience increased ROS when zinc limited. This is allowing
us to use the power of yeast genetics to identify the source of
oxidants in yeast and this may inform us about other processes
that generate ROS in zinc-limited mammalian cells. Further-
more, we have found a direct regulatory link in yeast between
antioxidant gene expression and zinc deficiency through the
Zap1 transcription factor. Zap1 mediates a proactive rather
than a reactive response to oxidative stress by up-regulating
antioxidants in response to low zinc. It will be interesting to
determine if mammalian cells also link regulation of anti-
oxidant enzymes directly to zinc status.
Acknowledgements
Work in the author’s lab on zinc homeostasis and oxidative
stress is funded by NIH grants GM056285 and GM093303.
The author apologizes to the many colleagues whose work was
not cited due the brief format of this minireview.
References
1 C. Andreini, L. Banci, I. Bertini and A. Rosato, Counting the zinc-
proteins encoded in the human genome, J. Proteome Res., 2006,
5(1), 196–201.
2 K. H. Brown, J. A. Rivera, Z. Bhutta, R. S. Gibson, J. C. King and
B. Lonnerdal, et al., International Zinc Nutrition Consultative
Group (IZiNCG) technical document #1. Assessment of the risk of
zinc deficiency in populations and options for its control, Food
Nutr. Bull., 2004, 25(1 Suppl 2), S99–203.
3 N. Leone, D. Courbon, P. Ducimetiere and M. Zureik, Zinc,
copper, and magnesium and risks for all-cause, cancer, and
cardiovascular mortality, Epidemiology, 2006, 17(3), 308–14.
4 T. Wu, C. T. Sempos, J. L. Freudenheim, P. Muti and E. Smit,
Serum iron, copper and zinc concentrations and risk of cancer
mortality in US adults, Ann. Epidemiol., 2004, 14(3), 195–201.
5 T. M. Bray and W. J. Bettger, The physiological role of zinc as an
antioxidant, Free Radical Biol. Med., 1990, 8(3), 281–91.
6 S. R. Powell, The antioxidant properties of zinc, J. Nutr., 2000,
130(5S Suppl), 1447S–54S.
7 P. I. Oteiza, M. S. Clegg, M. P. Zago and C. L. Keen, Zinc
deficiency induces oxidative stress and AP-1 activation in 3T3 cells,
Free Radical Biol. Med., 2000, 28(7), 1091–9.
8 B. Halliwell and M. Whiteman, Measuring reactive species and
oxidative damage in vivo and in cell culture: how should you do it
and what do the results mean?, Br. J. Pharmacol., 2004, 142(2),
231–55.
9 E. Ho and B. N. Ames, Low intracellular zinc induces oxidative
DNA damage, disrupts p53, Nfkappa B, and AP1 DNA binding,
and affects DNA repair in a rat glioma cell line, Proc. Natl. Acad.
Sci. U. S. A., 2002, 99(26), 16770–5.
10 E. Ho, C. Courtemanche and B. N. Ames, Zinc deficiency induces
oxidative DNA damage and increases p53 expression in human
lung fibroblasts, J. Nutr., 2003, 133(8), 2543–8.
11 A. Kojima-Yuasa, K. Umeda, T. Ohkita, D. Opare Kennedy,
S. Nishiguchi and I. Matsui-Yuasa, Role of reactive oxygen species
in zinc deficiency-induced hepatic stellate cell activation, Free
Radical Biol. Med., 2005, 39(5), 631–40.
12 M. P. Zago, G. G. Mackenzie, A. M. Adamo, C. L. Keen and
P. I. Oteiza, Differential modulation of MAP kinases by zinc
deficiency in IMR-32 cells: role of H(2)O(2), Antioxid. Redox
Signaling, 2005, 7(11–12), 1773–82.
13 M. Yan, Y. Song, C. P. Wong, K. Hardin and E. Ho, Zinc
deficiency alters DNA damage response genes in normal human
prostate epithelial cells, J. Nutr., 2008, 138(4), 667–73.
14 L. Aimo, G. N. Cherr and P. I. Oteiza, Low extracellular zinc
increases neuronal oxidant production through nadph oxidase and
nitric oxide synthase activation, Free Radical Biol. Med., 2010,
48(12), 1577–87.
1128 Metallomics, 2011, 3, 1124–1129
15 J. F. Sullivan, M. M. Jetton, H. K. Hahn and R. E. Burch,
Enhanced lipid peroxidation in liver microsomes of zinc-deficient
rats, Am. J. Clin. Nutr., 1980, 33(1), 51–6.
16 T. M. Bray, S. Kubow and W. J. Bettger, Effect of dietary zinc on
endogenous free radical production in rat lung microsomes,
J. Nutr., 1986, 116(6), 1054–60.
17 J. D. Hammermueller, T. M. Bray and W. J. Bettger, Effect of zinc
and copper deficiency on microsomal NADPH-dependent active
oxygen generation in rat lung and liver, J. Nutr., 1987, 117(5),
894–901.
18 P. I. Oteiza, K. L. Olin, C. G. Fraga and C. L. Keen, Zinc
deficiency causes oxidative damage to proteins, lipids and DNA
in rat testes, J. Nutr., 1995, 125(4), 823–9.
19 A. A. Shaheen and A. A. el-Fattah, Effect of dietary zinc onlipid
peroxidation, glutathione, protein thiols levels and superoxide
Downloaded from https://academic.oup.com/metallomics/article/3/11/1124/6016256 by guest on 03 November 2021
dismutase activity in rat tissues, Int. J. Biochem. Cell Biol., 1995,
27(1), 89–95.
20 R. Canali, F. Vignolini, F. Nobili and E. Mengheri, Reduction of
oxidative stress and cytokine-induced neutrophil chemoattractant
(CINC) expression by red wine polyphenols in zinc deficiency
induced intestinal damage of rat, Free Radical Biol. Med., 2000,
28(11), 1661–70.
21 R. S. Bruno, Y. Song, S. W. Leonard, D. J. Mustacich,
A. W. Taylor and M. G. Traber, et al., Dietary zinc restriction
in rats alters antioxidant status and increases plasma F2 isopros-
tanes, J. Nutr. Biochem., 2007, 18(8), 509–18.
22 Y. Song, V. Elias, A. Loban, A. G. Scrimgeour and E. Ho, Marginal
zinc deficiency increases oxidative DNA damage in the prostate after
chronic exercise, Free Radical Biol. Med., 2010, 48(1), 82–8.
23 Y. Song, S. W. Leonard, M. G. Traber and E. Ho, Zinc deficiency
affects DNA damage, oxidative stress, antioxidant defenses, and
DNA repair in rats, J. Nutr., 2009, 139(9), 1626–31.
24 A. S. Prasad, F. W. Beck, B. Bao, J. T. Fitzgerald, D. C. Snell and
J. D. Steinberg, et al., Zinc supplementation decreases incidence of
infections in the elderly: effect of zinc on generation of cytokines
and oxidative stress, Am. J. Clin. Nutr., 2007, 85(3), 837–44.
25 Y. Song, C. S. Chung, R. S. Bruno, M. G. Traber, K. H. Brown
and J. C. King, et al., Dietary zinc restriction and repletion affects
DNA integrity in healthy men, Am. J. Clin. Nutr., 2009, 90(2),
321–8, PMCID: 2709309.
26 E. Ho, Zinc deficiency, DNA damage and cancer risk., J. Nutr.
Biochem., 2004, 15(10), 572–8.
27 B. N. Ames and P. Wakimoto, Are vitamin and mineral deficien-
cies a major cancer risk?, Nat. Rev. Cancer, 2002, 2(9), 694–704.
28 C. Y. Wu, A. J. Bird, D. R. Winge and D. J. Eide, Regulation of
the yeast TSA1 peroxiredoxin by ZAP1 is an adaptive response to the
oxidative stress of zinc deficiency, J. Biol. Chem., 2007, 282(4), 2184–95.
29 C. Y. Wu, S. Roje, F. J. Sandoval, A. J. Bird, D. R. Winge and
D. J. Eide, Repression of sulfate assimilation is an adaptive
response of yeast to the oxidative stress of zinc deficiency,
J. Biol. Chem., 2009, 284(40), 27544–56, PMCID: 2785683.
30 C. Y. Wu, J. Steffen and D. J. Eide, Cytosolic superoxide
dismutase (SOD1) is critical for tolerating the oxidative stress of zinc
deficiency in yeast, PLoS One, 2009, 4(9), e7061, PMCID: 2737632.
31 K. Jomova and M. Valko, Advances in metal-induced oxidative
stress and human disease, Toxicology, 2011, 283(2–3), 65–87.
32 W. Maret, Cellular zinc and redox states converge in the metallo-
thionein/thionein pair, J. Nutr., 2003, 133(5 Suppl 1), 1460S–2S.
33 W. Maret, Metallothionein redox biology in the cytoprotective and
cytotoxic functions of zinc, Exp. Gerontol., 2008, 43(5), 363–9.
34 M. P. Zago and P. I. Oteiza, The antioxidant properties of zinc:
interactions with iron and antioxidants, Free Radical Biol. Med.,
2001, 31(2), 266–74.
35 M. P. Zago, S. V. Verstraeten and P. I. Oteiza, Zinc in the
prevention of Fe2+ initiated lipid and protein oxidation, Biol.
Res., 2000, 33(2), 143–50.
36 H. J. Coyne, 3rd, S. Ciofi-Baffoni, L. Banci, I. Bertini, L. Zhang
and G. N. George, et al., The characterization and role of zinc
binding in yeast Cox4, J. Biol. Chem., 2007, 282(12), 8926–34.
37 A. Atkinson, O. Khalimonchuk, P. Smith, H. Sabic, D. Eide and
D. R. Winge, Mzm1influences a labile pool of mitochondrial zinc
important for respiratory function, J. Biol. Chem., 2010, 285(25),
19450–9, PMCID: 2885224.
38 A. R. Frand, J. W. Cuozzo and C. A. Kaiser, Pathways for protein
disulphide bond formation, Trends Cell Biol., 2000, 10(5), 203–10.
This journal is c The Royal Society of Chemistry 2011
39 B. P. Tu and J. S. Weissman, Oxidative protein folding in
eukaryotes: mechanisms and consequences, J. Cell Biol., 2004,
164(3), 341–6.
40 B. P. Tu and J. S. Weissman, The FAD- and O(2)-dependent
reaction cycle of Ero1-mediated oxidative protein folding in the
endoplasmic reticulum, Mol. Cell, 2002, 10(5), 983–94.
41 E. Gross, C. S. Sevier, N. Heldman, E. Vitu, M. Bentzur and
C. A. Kaiser, et al., Generating disulfides enzymatically: reaction
products and electron acceptors of the endoplasmic reticulum thiol
oxidase Ero1p, Proc. Natl. Acad. Sci. U. S. A., 2006, 103(2), 299–304.
42 J. W. Cuozzo and C. A. Kaiser, Competition between glutathione
and protein thiols for disulphide-bond formation, Nat. Cell Biol.,
1999, 1(3), 130–5.
43 C. M. Haynes, E. A. Titus and A. A. Cooper, Degradation of
misfolded proteins prevents ER-derived oxidative stress and cell
death, Mol. Cell, 2004, 15(5), 767–76.
44 L. Li and J. Kaplan, The yeast gene MSC2, a member of the cation
diffusion facilitator family, affects the cellular distribution of zinc,
J. Biol. Chem., 2000, 276, 5036–43.
45 C. D. Ellis, C. W. Macdiarmid and D. J. Eide, Heteromeric protein
complexes mediate zinc transport into the secretory pathway of
eukaryotic cells, J. Biol. Chem., 2005, 280(31), 28811–8.
46 C. D. Ellis, F. Wang, C. W. MacDiarmid, S. Clark, T. Lyons and
D. J. Eide, Zinc and the Msc2 zinc transporter protein are required
for endoplasmic reticulum function, J. Cell Biol., 2004, 166(3),
325–35, PMCID: 2172251.
47 W. Droge, Free radicals in the physiological control of cell
function, Physiol. Rev., 2002, 82(1), 47–95.
48 A. M. Brennan, S. W. Suh, S. J. Won, P. Narasimhan,
T. M. Kauppinen and H. Lee, et al., NADPH oxidase is the
primary source of superoxide induced by NMDA receptor activa-
tion, Nat. Neurosci., 2009, 12(7), 857–63, PMCID: 2746760.
49 P. Paoletti, A. M. Vergnano, B. Barbour and M. Casado, Zinc at
glutamatergic synapses, Neuroscience, 2009, 158(1), 126–36.
50 D. J. Eide, Zinc transporters and the cellular trafficking of zinc,
Biochim. Biophys. Acta, Mol. Cell Res., 2006, 1763(7), 711–22.
51 M. R. Bleackley and R. T. Macgillivray, Transition metal homeo-
stasis: from yeast to human disease, Biometals, 2011.
This journal is c The Royal Society of Chemistry 2011
52 R. A. Colvin, W. R. Holmes, C. P. Fontaine and W. Maret,
Cytosolic zinc buffering and muffling: their role in intracellular
zinc homeostasis, Metallomics, 2010, 2(5), 306–17.
53 L. A. Lichten and R. J. Cousins, Mammalian zinc transporters:
nutritional and physiologic regulation, Annu. Rev. Nutr., 2009, 29,
153–76.
54 D. J. Eide, Homeostatic and Adaptive Responses to Zinc
Deficiency in Saccharomyces cerevisiae, J. Biol. Chem., 2009,
284(28), 18565–9, PMCID: 2707215.
55 D. S. Yuan, Zinc-regulated genes in Saccharomyces cerevisiae
revealed by transposon tagging, Genetics, 2000, 156(1), 45–58.
56 T. J. Lyons, A. P. Gasch, L. A. Gaither, D. Botstein, P. O. Brown
and D. J. Eide, Genome-wide characterization of the Zap1p zinc-
responsive regulon in yeast, Proc. Natl. Acad. Sci. U. S. A., 2000,
97(14), 7957–62, PMCID: 16652.
Downloaded from https://academic.oup.com/metallomics/article/3/11/1124/6016256 by guest on 03 November 2021
57 C. Y. Wu, A. J. Bird, L. M. Chung, M. A. Newton, D. R. Winge
and D. J. Eide, Differential control of Zap1-regulated genes in
response to zinc deficiency in Saccharomyces cerevisiae, BMC
Genomics, 2008, 9, 370.
58 R. De. Nicola, L. A. Hazelwood, E. A. De. Hulster, M. C. Walsh,
T. A. Knijnenburg and M. J. Reinders, et al., Physiological and
transcriptional responses of Saccharomyces cerevisiae to zinc
limitation in chemostat cultures, Appl. Environ. Microbiol., 2007,
73(23), 7680–92.
59 V. J. Higgins, P. J. Rogers and I. W. Dawes, Application of
genome-wide expression analysis to identify molecular markers
useful in monitoring industrial fermentations, Appl. Environ.
Microbiol., 2003, 69(12), 7535–40.
60 P. D. Bandara, J. A. Flattery-O’Brien, C. M. Grant and
I. W. Dawes, Involvement of the Saccharomyces cerevisiae
UTH1 gene in the oxidative-stress response, Curr. Genet., 1998,
34(4), 259–68.
61 D. Thomas and Y. Surdin-Kerjan, Metabolism of sulfur amino
acids in Saccharomyces cerevisiae, Microbiol. Mol. Biol. Rev., 1997,
61(4), 503–32.
62 P. Kaiser, N. Y. Su, J. L. Yen, I. Ouni and K. Flick, The yeast
ubiquitin ligase SCFMet30: connecting environmental and intra-
cellular conditions to cell division, Cell Div., 2006, 1, 16.
Metallomics, 2011, 3, 1124–1129 1129