TRPLSC 2480 No.

of Pages 3

Trends in
Plant Science
Spotlight
The two faces of DJ-1D [2]. The detoxification system initiates with
the spontaneous conversion of MG into a
homologs is the presence of two DJ-1/
PfpI domains in most of the plants’ GLYIII
proteins hemithioacetal utilizing reduced GSH as members [7,8].
1 a cofactor. Of the two enzymes, GLY1
Priya Gambhir, then mediates the conversion of this In contrast to the sole function of the GSH-
Arun Kumar Sharma, 1 and hemithioacetal to S-lactoylglutathione, an- dependent glyoxalase system in MG de-
Rahul Kumar 2,* notated as the rate-limiting step of this toxification, GLYIII proteins also exhibit
pathway. Lastly, S-lactoylglutathione acts chaperone and protease activities and
Despite the documented bi- as a substrate for the GLYII enzyme to con- serve as dicarbonyl degraders. Recently,
enzymatic mode of methylglyoxal vert into a nontoxic compound, D-lactate, a new deglycase activity was assigned to
detoxification, the single-step ca- recycling GSH back into the antioxidant human DJ-1 proteins. A Parkinsonism-
talysis of methylglyoxal by DJ-1/ pool for further metabolic usage [2]. Follow- associated protein, DJ-1/Park7, was re-
Pfp-I domain containing proteins ing its discovery in animals, a shorter and ported to repair MG-generated glycations
has been in the limelight. Prasad less energy-demanding route of MG con- in proteins, specifically at cysteine, lysine,
et al. recently discovered another version mediated by novel GLYIII proteins and arginine residues and in DNA and
functional facet of these moon- has also been recently identified in plants RNA molecules at guanine residues
lighting proteins: the deglycase [6]. The GLYIII proteins encompass the [9,10]. To simplify, GLYIII executes its
DJ-1/PfpI domains and show a lower deglycase activity by first recruiting its
potential of DJ-1D to repair the gly-
glyoxalase activity than GLY1 enzymes. chaperone role to interact with the non-
cated DNA, RNA, and proteins in
DJ-1-mediated MG detoxification is a native glycated proteins to gain access to
plants. GSH- and metal-ion-independent process partially buried glycated sites. Then,
[7]. One striking structural feature that dif- GLYIII proteins use their glyoxalase 1 and
ferentiates plant GLYIII from non-plant glyoxalase 2 competence to catalyze the

The production of methylglyoxal (MG), a

highly reactive di-carbonyl compound, is
often tightly bound to cellular respiration;
therefore, its synthesis is inevitable in all
living cells [1,2]. After its initial discovery in
animal tissues as a potent growth suppres-
sant, MG has been found to induce irre-
versible change in nucleic acids, lipids,
and protein substrates to yield a heteroge-
nous group of molecules, collectively called
advanced glycation end products (AGEs)
[3]. The knowledge of the existence of MG
later transcended to plants and a consider-
able amount of evidence shows its
overaccumulation in plants under hostile Trends in Plant Science
growth conditions [4,5]. Although multiple
Figure 1. Overview of glutathione (GSH)-dependent and -independent glyoxalase-mediated
metabolic reactions can prevent MG methylglyoxal (MG) detoxification systems. MG is primarily synthesized from triosephosphate sugars
overaccumulation, the glyoxalase detoxi- dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (GAP) during glycolysis through
fication system, first discovered in 1913 nonenzymatic reactions. As a potent reactive oxygen species (ROS) intensifier, enhanced production of MG at
the time of stressful conditions triggers glycation in cysteine, arginine, and lysine amino acid residues in
by Dakin and Dudley, represents the pri-
proteins as well as guanosine and deoxyguanosine residues in nucleic acid. In order to maintain MG
mary pathway of its containment in living homeostasis, all living cells are equipped with the machinery to detoxify it (i.e., glyoxalase detoxification
cells. This bi-enzymic, glutathione (GSH)- system). The conventional glyoxalase mechanism is bi-enzymatic and GSH-dependent, using GLYI and GLYII
dependent, and energy-intensive catalytic enzymes to convert MG to a nontoxic compound D-lactate. However, the recently characterized MG
detoxification process is catalyzed by the DJ-1/Pfp1 (GLYIII) enzyme in a single step and in a GSH-
module encompassing the conventional independent fashion. Apart from harboring methylglyoxalase activity, DJ-1 proteins have been shown to
GLYI and GLYII enzymes converts MG to possess a strong deglycase activity, by virtue of which these moonlighting proteins can repair the glycation of
a nontoxic compound, D-lactate (Figure 1) amino acid and guanosine residues in proteins and nucleic acids, respectively.

Trends in Plant Science, Month 2023, Vol. xx, No. xx 1


conversion of hemithioacetals into thioesters
for cysteine deglycation. Eventually, it leads
to the cleavage of amide bonds to assist
lysine/arginine deglycation, using their
amidase/peptidase activities. This remark-
able moonlighting function of GLYIII
proteins was also recently demonstrated
in the plant kingdom. In a recent study,
Prasad et al. highlighted the sequence
similarity of Arabidopsis thaliana
(arabidopsis) DJ-1D with human DJ-1/
PARK-7 and Saccharomyces cerevisiae
Hsp31 (ScHsp31) proteins, thereby con-
templating similar physiological functions
[11]. Although the authors reported that
each DJ-1/PfpI domain of AtDJ-1D could in-
dependently detoxify MG, the full-length
double-domain protein displayed higher
MG scavenging potential. The authors
reported that mutations in the amino acid
residues of the core catalytic triad of
both domains could only desolate the
methylglyoxalase activity. It is well-known
that MG concentration positively correlates
to the glycation state of macromolecules
inside the cell. Although both GSH-
dependent and -independent systems are
activated in response to dicarbonyl intensifi-
cation in plants, the GLYIII-mediated detoxi-
fication mechanism appears to be highly
advantageous as it does not rely on the al-
ready compromised GSH pool under
stressful environments [5,6]. Moreover, the
robustness of AtDJ-1D protein in mitigating
responses against multiple endogenous
and exogenous stresses could be attributed
to its efficient repair mechanism to revert
dicarbonyl-mediated glycation end-
products of nucleic acids and proteins into
their native functional forms through its
deglycase activity. The authors showed
that the two domains of AtDJ-1D could
independently reduce MG-induced AGE
formation; however, a mutation in both do-
mains significantly abolished its deglycase
ability. Prasad et al. further maneuvered
several in vivo and in vitro chaperone as-
says of AtDJ-1D in S. cerevisiae using sev-
eral substrates, such as rhodanese and
human SOD1, and confirmed its functional
2 Trends in Plant Science, Month 2023, Vol. xx, No. xx
relevance in maintaining proteostasis in
plants. Interestingly, the repair mechanism
endeavored by AtDJ-1D is not confined to
protein glycation products only, as it rectified
the MG-induced glycation errors in guano-
sine and deoxyguanosine residues in DNA
and RNA in order to prevent the formation
of cytotoxic aminocarbinols and cyclic
imidazopurines, one of the most potent
AGEs. Since much of our understanding of
the plant DJ-1 proteins have come from
their previously reported counterparts in
human and yeast, it was fascinating to see
whether these homologs inhabit the same
cellular compartment or not. To find this,
the authors fused AtDJ-1D with GFP and
then mapped its cellular localization under
normal and stressful conditions. Human
DJ-1/PARK-7 protein localizes specifically
to nucleus and mitochondria [10]. In normal
growth conditions, AtDJ-1D localized to
both mitochondria and cytoplasm. How-
ever, in response to oxidative stress, the
protein localization shifted to mitochondria
and nucleus, a profile similar to ScHsp31
but different from its human isoform [12].
To investigate the role of this protein in
planta, Prasad et al. generated transgenic
arabidopsis and Nicotiana plants overex-
pressing AtDJ-1D. The transgenic plants ex-
hibited higher chlorophyll retention, lower
reactive oxygen species (ROS) accumula-
tion, and decreased MDA levels in conjunc-
tion with enhanced tolerance to multiple
stresses, including oxidative, salinity, and
osmotic stresses, than their wild-type
counterparts. The results perfectly reso-
nate with the previously documented
function of GLYIII proteins in imparting
stress tolerance against multiple abiotic
stresses [5]. Intriguingly, Prasad et al.
failed to generate arabidopsis DJ-1D
knock-down plants, even after repeated
attempts, implicating essential require-
ment of this gene in overall plant growth
and development.
The proposed model in Figure 1 displays
the two modes of MG detoxification pro- Declaration of interests
cesses present in living systems and how No interests are declared.
Trends in Plant Science
the DJ-1D driven mechanism is superior
to the GLYI-GLYII pathway in terms of
maintaining MG homeostasis but also by
rectifying MG-induced protein and nucleic
acid glycation.
Conclusions
To summarize, Prasad et al. conclusively
demonstrate a novel glycation repair
mechanism in plants involving DJ-1/
GLYIII proteins, which serve as mainte-
nance and repair hotspots of nucleic acids
and proteins in response to dicarbonyl
stresses. The functional characterization of
AtDJ-1D, in planta, for its phenomenal
methylglyoxalase and deglycase properties,
led to superior growth of transgenic plants,
further spotlighting the potential application
of this gene for engineering tolerance
against multiple stresses and improving
the overall yield of the plant. Although,
GLYIII members constitute a small subfam-
ily in plants, so far not all members have
been investigated for their functions in de-
tail. Considering their localization to differ-
ent cellular organelles and the improved
performance of DJ-1 homologs overex-
pressing plants over their wild-type con-
trols under multiple abiotic stresses in
tomato, tobacco, and arabidopsis, it will
be interesting to uncover the complete
range of their roles and delineate underlying
molecular mechanisms in relations to
plant’s fitness and adaptation to various
stress environments.
Acknowledgments
R.K. acknowledges the DBT (BT/PR31630/AGIII/103/
1119/2019), SERB (CRG/2018/001033), and MHRD-
IoE [(RC1-20-018) and (F11/9/2019-U3(A))] and DBT-
BUILDER for funding the laboratory. A.K.S. acknowl-
edges DBT grant BT/PR6983/PBD/16/1007/2012. The
authors acknowledge the Department of Science and
Technology, India, for the Purse Grant. The SAP Grant
of the University Grants Commission and FIST grant of
DST, India, to the Department of Plant and Molecular
Biology, UDSC, and the Department of Plant Sciences,
UoH, for infrastructure support are also acknowledged.
Trends in Plant Science
1 3. Bechtold, U. et al. (2009) Quantitative measurement of
Department of Plant Molecular Biology, University of Delhi
South Campus, New Delhi 110021, India
2 glycation in Arabidopsis leaves. Plant J. 59, 661–671
Department of Plant Sciences, School of Life Sciences,
University of Hyderabad, Hyderabad 500046, India
*Correspondence:
rksl@uohyd.ac.in (R. Kumar).
https://doi.org/10.1016/j.tplants.2023.06.005
© 2023 Elsevier Ltd. All rights reserved.
References
1. Takagi, D. et al. (2014) The Calvin cycle inevitably produces
sugar-derived reactive carbonyl methylglyoxal during photosyn-
thesis: a potential cause of plant diabetes. Plant Cell Physiol. 55,
333–340
2. Singla-Pareek, S.L. et al. (2020) Reassessing plant glyoxalases:
large family and expanding functions. New Phytol. 227,
714–721
10. Richarme, G. et al. (2015) Parkinsonism-associated protein
specific biomarkers for protein oxidation, nitration and DJ-1/Park7 is a major protein deglycase that repairs
methylglyoxal- and glyoxal-glycated cysteine, arginine,
4. Gupta, B.K. et al. (2018) Manipulation of glyoxalase path- and lysine residues. J. Biol. Chem. 290, 1885–1897
way confers tolerance to multiple stresses in rice. Plant 11. Prasad, M. et al. (2022) Double DJ-1 domain containing
Cell Environ. 41, 1186–1200 Arabidopsis DJ-1D is a robust macromolecule deglycase.
5. Gambhir, P. et al. (2023) A glutathione-independent DJ-1/ New Phytol. 236, 1061–1074
PfpI domain-containing tomato glyoxalaseIII2, SlGLYIII2, 12. Bankapalli, K. et al. (2015) Robust glyoxalase activity of Hsp31,
confers enhanced tolerance under salt and osmotic a ThiJ/DJ-1/PfpI family member protein, is critical for oxidative
stresses. Plant Cell Environ. 46, 518–548 stress resistance in Saccharomyces cerevisiae. J. Biol. Chem.
6. Ghosh, A. et al. (2016) Presence of unique glyoxalase III 290, 26491–26507
proteins in plants indicates the existence of shorter route
for methylglyoxal detoxification. Sci. Rep. 6, 18358
7. Kumar, B. et al. (2021) Tracing the evolution of plant
glyoxalase III enzymes for structural and functional
divergence. Antioxid. (Basel) 10, 648
8. Kwon, K. et al. (2013) Novel glyoxalases from Arabidopsis
thaliana. FEBS J. 280, 3328–3339
9. Richarme, G. et al. (2017) Guanine glycation repair by
DJ-1/Park7 and its bacterial homologs. Science 357,
208–211
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