Characteristic of Glutamate Cysteine Ligase Gene and its response to the salinity and temperature stress in Chlamydomonas sp. ICE-L from Antarctica
Chlamydomonas sp. ICE-L, an Antarctic ice
alga, has high tolerance ability to freezing and salinity. Glutathione
(GSH) is an important small antioxidative molecule in the growth and
stress responses of plants including algae. We cloned a full-length cDNA
encoding glutamate cysteine ligase (ICE-LGCL), the key enzyme of GSH
synthesis, from Chlamydomonas sp. ICE-L by RT-PCR and rapid
amplification of cDNA ends technique (RACE). The cDNA has 2199 bp
nucleotides with an open reading frame (ORF) of 1452 bp encoding a
polypeptide of 453 amino acids. BLASTP algorithm results showed that
ICE-LGCL shared 51%-70% amino acid sequence identity with the reported
GCLs and shared the highest identity with Chlamydomonas reinhardtii.
We also successfully made ICE-LGCL protein express in E. coli BL21. The
optimum expression conditions are the induced reagent IPTG of 0.2
mmol/L, temperature of 37 °C, and the induced time of 4 h. The
expression patterns of ICE-LGCL in mRNA by real-time PCR analysis showed
that ICE-LGCL expressed under the different temperature and salinity
challenges. Low temperature and low salinities stimulated the
accumulation of ICE-LGCL mRNA in ICE-L cells. These results indicate
that GCL might play an important role in Antarctic Chlamydomonas sp. ICE-L.
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- Anderson ME, Powrie F, Puri RJ, Meister A (1985). Glutathione
monoethylester: preparation uptake by tissues and conversion
to glutathione.
Arch Biochem Biophys 239: 538-548.
- Arteel GE, Sies H (2001). The biochemistry of selenium and the
glutathione system. Environ Toxicol Phar 10: 153-158.
- Copley SD, Dhillon JK (2002). Lateral gene transfer and parallel
evolution in the history of glutathione biosynthesis genes.
Genome Biol 3: 1-16.
- Dhankher OP, Li Y, Rosen BP, Shi J, Salt D, Senecoff JF, Sashti
NA, Meagher RB (2002). Engineering tolerance and hyper
accumulation of arsenic in plants by combining arsenate
reductase and gamma-glutamylcysteine synthetase expression.
Nat Biotechnol 20:
1140-1145.
- Dickinson DA, Forman HJ (2002). Glutathione in defense and
signaling lessons from a small thiol.
Ann NY Acad Sci 973:
488-504.
- Dickinson DA, Levonen AL, Moellering DR, Arnold EK, Zhang
H, Darley-Usmar VM, Forman HJ (2004). Human glutamate
cysteine ligase gene regulation through the electrophile
response element. Free Radical Bio Med 37: 1152-1159.
- Ding Y, Miao JL, Wang QF, Li GY (2005). Effect of Cd on GSH and
GSH-related enzymes of
Chlamydomonas
sp. ICE-L existing in
Antarctic ice. J Environ Sci-China 17: 667-671.
- Ding Y, Miao JL, Wang QF, Li GY (2007). Influence of temperature on
glutathione level and glutathione-related enzyme activities of
Antarctic ice microalgae
Chlamydomonas
sp. ICE-L.
Chinese
Journal of Polar Research 18: 161-167.
- Ding Y, Liu Y, Jian JC, Wu ZH, Miao JL (2012). Molecular cloning
and expression analysis of glutathione reductase gene in
Chlamydomonas
sp. ICE-L from Antarctica.
Mar Genom 5:
59-64.
- Eggler AL, Gay KA, Mesecar AD (2008). Molecular mechanisms
of natural products in chemoprevention: induction of
cytoprotective enzymes by Nrf2. Mol Nutr Food Res 52 Suppl
1: S84-94.
- Gomez LD, Vanacker H, Buchner P, Noctor G, Foyer CH (2004).
Intercellular distribution of glutathione synthesis in maize
leaves and its response to short-term chilling. Plant Physiol
134: 1662-1671.
- Griffith OW (1999). Biologic and pharmacologic regulation of
mammalian glutathione synthesis. Free Radical Bio Med 27:
922-935.
- Hell R, Bergmann L (1990). γ-Glutamylcysteine synthetase in higher
plants: catalytic properties and subcellular localization.
Planta,
180: 603-612.
- Hiroyuki A, Yoshihiro S, Hitoshi S (2005). Cloning, biochemical
and phylogenetic characterizations of γ-glutamylcysteine
synthetase from
Anabaena sp
. PCC 7120. Plant Cell Physiol 46:
557-562.
- Iwamoto K, Shiraiwa Y (2005). Salt-regulated mannitol metabolism
in algae. Mar Biotechnol 7: 407-415.
- Kaminaka H, Morita S, Nakajima M, Masumura T, Tanaka K (1998).
Gene cloning and expression of cytosolic glutathione reductase
in rice (
Oryza sativa
). Plant Cell Physiol 39: 1269-1280.
- Kan GF, Zheng Z, Jiang YH, Miao JL, Zhang BT, Li GY (2006). Salt
resistance of Antarctic ice microalga
Chlamydomonas
sp.
ICE-L. Journal of Fishery Sciences of China 13: 73-78.
- Kocsy G, Brunner M, Ruegsegger A, Stamp P, Brunold C (1996).
Glutathione synthesis in maize genotypes with different
sensitivities to chilling. Planta 198: 365-370.
- Kocsy G, Owttrim G, Brander K, Brunold C (1997). Effect of chilling
on the diurnal rhythm of enzymes involved in protection
against oxidative stress in a chilling-tolerant and a chilling-
sensitive maize genotype. Physiol Plantarum 99: 249-254.
- Kocsy G, Szalai G, Vagujfalvi A, Stehli L, Orosz G, Galiba G (2000).
Genetic study of glutathione accumulation during cold
hardening in wheat. Planta 210: 295-301.
- Lu SC (2000). Regulation of glutathione synthesis. Curr Top Cell
Regul 36: 95-116.
- May MJ, Vernoux T, Leaver C, Van MM, Inze D (1998a). Glutathione
homeostasis in plants: implications for environmental sensing
and plant development. J Exp Bot 49: 649-667.
- May MJ, Vernoux T, Sánchez-Fernández R, Van MM, Inze D
(1998b). Evidence for posttranscriptional activation of gamma-
glutamylcysteine synthetase during plant stress responses. P
Natl Acad Sci USA 95: 12049-12054.
- Mittova V, Theodoulou FL, Kiddle G, Gomez L, Volokita M, Tal M,
Foyer CH, Guy M (2003). Coordinate induction of glutathione
biosynthesis and glutathione-metabolizing enzymes is
correlated with salt tolerance in tomato. FEBS Lett 554: 417-
421.
- Mock T, Thomas DN (2005). Recent advances in sea-ice microbiology.
Environ Microbiol 7: 605-619.
- Noctor G, Arisi A, Jouanin L, Kunert K, Rennenberg H, Foyer C
(1998). Glutathione: biosynthesis, metabolism and relationship
to stress tolerance explored in transformed plants. J Exp Bot
49: 623-647.
- Ogawa K (2005). Glutathione-associated regulation of plant growth
and stress responses.
Antioxid Redox Sign 7: 973-981.
- Ohtake Y, Yabuuchi S (1991). Molecular cloning of the
γ-glutamylcysteine synthetase gene of
Saccharomyces
cerevisiae
. Yeast 7: 953-961.
- Provasoli L (1968). Media and prospects for the cultivation of marine
algae. In: Culture and collections of algae (Watanabe A, Hattori
R, editors), 63-95. Proceedings of the US-Japan Conference,
Hakone, Japan.
- Reed DJ, Fariss MW (1984). Glutathione depletion and susceptibility.
Pharmacol Rev 36 (2 Suppl): 25S-33S.
- Reid LL, Botta D, Lu Y, Gallagher EP, Kavanagh TJ (1997). Molecular
cloning and sequencing of the cDNA encoding the catalytic
subunit of mouse glutamate-cysteine ligase. Biochim Biophys
Acta 1352: 233-237.
- Reisinger S, Schiavon M, Terry N, Pilon-Smits EA (2008). Heavy
metal tolerance and accumulation in Indian mustard (
Brassica
juncea
L.) expressing bacterial gamma-glutamylcysteine
synthetase or glutathione synthetase. Int J Phytoremediat 10:
440-54.
- Ruiz JM, Blumwald E (2002). Salinity-induced glutathione synthesis
in
Brassica napus
. Planta 214: 965-969.
- Sandrini JZ, Laurino J, Hatanaka T, Monserrat JM, Marins LF (2006).
cDNA cloning and expression analysis of the catalytic subunit
of glutamate cysteine ligase gene in an annelid polychaete
after cadmium exposure: A potential tool for pollution
biomonitoring. Comp Biochem Physiol
C 143: 410-415.
- Schäfer HJ, Greiner S, Rausch T, Haag-Kerwer A (1997). In seedlings of
the heavy metal accumulator
Brassica juncea
Cu
2+
differentially
affects transcript amounts for gamma-glutamylcysteine
synthetase (gamma-ECS) and metallothionein (MT2). FEBS
Lett 404: 216-220.
- Sharma SK, Banyal HS (2009). γ-glutamylcysteine synthetase from
Plasmodium berghei
. Parasitol Int 58: 145-153.
- Sunagawa S, Choi J, Forman HJ, Medina M (2008). Hyperthermic
stress-induced increase in the expression of glutamate-cysteine
ligase and glutathione levels in the symbiotic sea anemone
Aiptasia pallida
. Comp Biochem Physiol
B 151: 133-138.
- Thomas ND, Dieckmann GS (2002). Antarctic sea ice - a habitat for
extremophiles. Science 295: 641-644.
- Thomashow MF (2001). So what’s new in the field of plant cold
acclimation? Lots! Plant Physiol 125: 89-93.
- Wang HY, Zhu F, Huang YC, Ding Y, Jian JC, Wu ZH (2017).
Construction of glutathione peroxidase (GPx) DNA vaccine
and its protective efficiency on the orange-spotted grouper
(
Epinephelus coioides
) challenged with
Vibrio harveyi
. Fish
Shellfish Immun 60: 529-536.
- Wu J, Zhao Z, An L, Liu Y, Xu S, Gao D, Zhang Y (2008). Inhibition of
glutathione synthesis decreases chilling tolerance in
Chorispora
bungeana
callus. Cryobiology 57: 9-17.
- Wu JM, Qu T, Chen SY, Zhao ZG, An LZ (2009). Molecular cloning
and characterization of a
γ
-glutamylcysteine synthetase gene
from
Chorispora bungeana
. Protoplasma 235: 27-36.
- Yin J, Li G, Ren X, Herrler G (2007). Select what you need: a
comparative evaluation of the advantages and limitations
of frequently used expression systems for foreign genes. J
Biotechnol 127: 335-347.
- Zhang BY, Fang Y, Wang GC, Peng G (2010). Cloning and quantitative
analysis of carbonic anhydrase gene from
Porphyra yezoensis
. J
Phycol 46: 290-296.
- Zheng Z, Miao JL, Kan GF, Jin Q, Ding Y, Liu FM, Wang SQ, Wang
YB (2010). Effect of high salinity on cell growth and protein
production of Antarctic ice microalgae
Chlamydomonas sp
.
ICE-L. Chinese Journal of Polar Research 21: 81-90.
- Zhu YL, PilonSmits EAH, Tarun AS, Weber SU, Jouanin L,
Terry N (1999). Cadmium tolerance and accumulation in
Indian mustard is enhanced by overexpressing gamma-
glutamylcysteine synthetase. Plant Physiol 121: 1169-1177.