Identification of a developmentally and hormonally regulated Delta-Class glutathione S-transferase in rice moth Corcyra cephalonica

Department of Animal Sciences, University of Hyderabad, India.
Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology (Impact Factor: 1.55). 02/2010; 156(1):33-9. DOI: 10.1016/j.cbpb.2010.01.016
Source: PubMed


Glutathione S-transferases (GSTs) are a large family of multifunctional enzymes, known for their role in cellular detoxification. Here we report a cytosolic GST with optimal activity at alkaline pH (8.3) from the visceral fat body of late-last instar (LLI) larvae of a lepidopteran insect rice moth Corcyra cephalonica. All previously known GSTs are active between pH 6.0 to 6.5. Purification and characterization revealed the Corcyra cephalonica GST (CcGST) as a 23-kDa protein. HPLC and 2D analysis showed a single isoform of the protein in the LLI visceral fat body. Degenerate primer based method identified a 701-nucleotide cDNA and the longest open reading frame contained 216 amino acids. Multiple sequence and structural alignment showed close similarity with delta-class GSTs. CcGST is present mainly in the fat body with highest activity at the late-last instar larval stage. Juvenile hormone (JH) negatively inhibits the CcGST activity both ex vivo and in vivo. We speculate that high expression and activity of CcGST in the fat body of the late-last instar larvae, when endogenous JH titer is low may have role in the insect post-embryonic development unrelated to their previously known function.

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    • "GSTs from Anopheles gambiae and cutworms Spodoptera litura are regulated variously in different developmental stages (Strode et al., 2006; Huang et al., 2011). A delta GST identified from Corcyra cephalonica has been proposed to be involved in development, according to its expression profile and hormone-regulated activity (Gullipalli et al., 2010). The expression of the sigma and delta classes of GST in Bombyx mori is regulated by a juvenile hormone analog (JHA) and 20-hydroxyecdysone (Zou et al., 2011). "
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    ABSTRACT: The juvenile hormone titer during the last instar is a crucial determining switch for metamorphosis in lepidopterans. We previously observed that the induction of glutathione S-transferase (GST) activity by a juvenile hormone analog (JHA) is reversely proportional to the occurrence of JHA-induced supernumerary instars following topical application of JHA to Spodoptera litura during the last instar period. In this paper, at least five JHA-induced GSTs were purified by glutathione-affinity chromatography, and one was further isolated for determination of the N-terminal sequence. The corresponding cDNA was cloned and named SIGST1 (GenBank accession no. AY506545). SIGST1 was classified into the epsilon class of GSTs by phylogenetic analysis. Northern blot analyses further showed that the SIGST1 in fat bodies can be induced by JHA, particularly in 0-clay-old sixth-instar larvae, in which induction was much higher than that in 1- and 2-day-old sixth-instar larvae. To our knowledge, this is the first JHA-inducible GST to have been identified, and we believe that it will provide new insight into the role of GST in insect development, particularly during metamorphosis.
    Journal of Asia-Pacific Entomology 09/2014; 17(3):363–368. DOI:10.1016/j.aspen.2014.02.005 · 0.95 Impact Factor
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    • "In this context, it was reported that CYP genes are upregulated in presence of xenobiotic or insecticidal substances through both constitutive overexpression and induction mechanisms (Poupardin et al. 2010, Liu et al. 2011). Glutathione-S-transferase (GST): GSTs play a central role in the detoxification of both endogenous and xenobiotic compounds and they are also involved in intracellular transport, biosynthesis of hormones and protection against oxidative stress (Gullipalli et al. 2010, Huang et al. 2011). In insects, there are two major classes of GSTs classified according to their location within the cell: microsomal and cytosolic. "
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    ABSTRACT: Anthonomus grandis Boheman is a key pest in cotton crops in the New World. Its larval stage develops within the flower bud using it as food and as protection against its predators. This behavior limits the effectiveness of its control using conventional insecticide applications and biocontrol techniques. In spite of its importance, little is known about its genome sequence and, more important, its specific expression in key organs like the midgut. Total mRNA isolated from larval midguts was used for pyrosequencing. Sequence reads were assembled and annotated to generate a unigene data set. In total, 400,000 reads from A. grandis midgut with an average length of 237 bp were assembled and combined into 20,915 contigs. The assembled reads fell into 6,621 genes models. BlastX search using the NCBI-NR database showed that 3,006 unigenes had significant matches to known sequences. Gene Ontology (GO) mapping analysis evidenced that A. grandis is able to transcripts coding for proteins involved in catalytic processing of macromolecules that allows its adaptation to very different feeding source scenarios. Furthermore, transcripts encoding for proteins involved in detoxification mechanisms such as p450 genes, glutathione-S-transferase, and carboxylesterases are also expressed. This is the first report of a transcriptomic study in A. grandis and the largest set of sequence data reported for this species. These data are valuable resources to expand the knowledge of this insect group and could be used in the design of new control strategies based in molecular information. © The Author 2014. Published by Oxford University Press on behalf of the Entomological Society of America.
    Journal of Insect Science 07/2014; 14(1). DOI:10.1093/jisesa/ieu081 · 1.03 Impact Factor
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    • "Although most eukaryotic GSTs are known to have optimal pH ranging between 6.0 and 6.5 [31], some reports suggest that insect GSTs also have considerable activities at higher pH. For example, Corcyra cephalonica GST has an optimal pH 8.3 [4]. A recombinant GSTt1 expressed in E.coli from B. mori showed broadly optimal pH ranging between 4.0 and 9.0 [27]. "
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    ABSTRACT: Glutathione S-transferases (GSTs) play an important role in detoxification of xenobiotics in both prokaryotic and eukaryotic cells. In this study, four GSTs (LmGSTd1, LmGSTs5, LmGSTt1, and LmGSTu1) representing different classes were identified from the migratory locust, Locusta migratoria. These four proteins were heterologously expressed in Escherichia coli as soluble fusion proteins, purified by Ni(2+)-nitrilotriacetic acid agarose column and biochemically characterized. LmGSTd1, LmGSTs5, and LmGSTu1 showed high activities with 1-chloro-2, 4-dinitrobenzene (CDNB), detectable activity with p-nitro-benzyl chloride (p-NBC) and 1, 2-dichloro-4-nitrobenzene (DCNB), whereas LmGSTt1 showed high activity with p-NBC and detectable activity with CDNB. The optimal pH of the locust GSTs ranged between 7.0 to 9.0. Ethacrynic acid and reactive blue effectively inhibited all four GSTs. LmGSTs5 was most sensitive to heavy metals (Cu(2+) and Cd(2+)). The maximum expression of the four GSTs was observed in Malpighian tubules and fat bodies as evaluated by western blot. The nymph mortalities after carbaryl treatment increased by 28 and 12% after LmGSTs5 and LmGSTu1 were silenced, respectively. The nymph mortalities after malathion and chlorpyrifos treatments increased by 26 and 18% after LmGSTs5 and LmGSTu1 were silenced, respectively. These results suggest that sigma GSTs in L. migratoria play a significant role in carbaryl detoxification, whereas some of other GSTs may also involve in the detoxification of carbaryl and chlorpyrifos.
    PLoS ONE 03/2013; 8(3):e58410. DOI:10.1371/journal.pone.0058410 · 3.23 Impact Factor
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