Detection and functional characterization of sigma class GST in Phlebotomus argentipes and its role in stress tolerance and DDT resistance

Hassan, Faizan; Singh, Krishn Pratap; Ali, Vahab; Behera, Sachidananda; Shivam, Pushkar; Das, Pradeep; Dinesh, Diwakar Singh

doi:10.1038/s41598-019-56209-0

Cited by 27 publications

(15 citation statements)

References 66 publications

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“…For example, AccGSTS1 in the honey bee A. cerana cerana was upregulated in response to phoxim, cyhalothrin and acaricide treatment (Yan et al, 2013). The GSTS PargGSTσ in P. argentipes was upregulated in a DDT-resistant strain compared with the susceptible strain (Hassan et al, 2019). After treatment of T. castaneum with phoxim, lambda-cyhalothrin, dichlorvos and carbofuran in our study, TcGSTs6 was significantly induced.…”

Section: Discussionsupporting

confidence: 44%

“…In vitro peroxidase activity and 4-HNE activity assays revealed that TcGSTS6 had high activity toward cumene hydroperoxide and 4-HNE. The peroxidase activity of GSTs promotes the reduction of organic hydroperoxides and plays an important role in various insects (Parkes et al, 1993), such as D. melanogaster, P. argentipes and A. cerana cerana (Yan et al, 2013;Hassan et al, 2019). 4-HNE is a cytotoxic product of lipid peroxidation under oxidative stress (Ayala et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

“…AccGSTS1 in the Chinese honey bee Apis cerana cerana could remove H 2 O 2 , and Escherichia coli overexpressing AccGSTS1 showed longterm resistance to oxidative stress (Yan et al, 2013). The GSTS PargGSTσ in Phlebotomus argentipes conferred DDT resistance by enhancing oxidative stress tolerance (Hassan et al, 2019). Glyphosate exposure in the freshwater planarian Dugesia japonica induced messenger RNA (mRNA) transcription and enzymatic activity of a GSTS (Zhang et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

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Characterization of a sigma class GST (GSTS6) required for cellular detoxification and embryogenesis in Tribolium castaneum

Song

Zhong

et al. 2021

Insect Science

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The sigma glutathione S-transferases (GSTSs) are a class of cytosolic glutathione S transferases (GSTs) that play important roles in antioxidant defense in insects, but the mechanisms by which GSTSs contribute to antioxidant activity remain unclear. Here, we isolated a GSTS (GSTS6) from Tribolium castaneum and explored its function. Homology and phylogenetic analysis revealed that TcGSTS6 shared high identity with other evolutionarily conserved GSTSs. The recombinant TcGSTS6 protein had strong activity toward cumene hydroperoxide and 4-hydroxynonenal but low activity toward the universal substrate 1-chloro-2,4-dinitrobenzene. Exposure to various types of oxidative stress, including heat, cold, UV and pathogenic microbes, significantly induced TcGSTs6 expression, which indicates that it is involved in antioxidant defense. Knockdown TcGSTs6 by using RNA interference (RNAi) caused reduced antioxidant capacity, which was accomplished by cooperating with other antioxidant genes. Moreover, treatment with various insecticides such as phoxim, lambda-cyhalothrin, dichlorvos and carbofuran revealed that TcGSTS6 plays an important role in insecticide detoxification. The RNAi results showed that TcGSTS6 is essential for embryogenesis in T. castaneum. Our study elucidates the mechanism by which a GSTS contributes to antioxidant activity and enhances our understanding of the functional diversity of GSTSs in insects.

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Section: Discussionsupporting

confidence: 44%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterization of a sigma class GST (GSTS6) required for cellular detoxification and embryogenesis in Tribolium castaneum

Song

Zhong

et al. 2021

Insect Science

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“…Peroxidase activity is of particular importance because in contrast to Se-dependent GSH-peroxidase activity, GST peroxidase is Se independent and may having a role in cellular antioxidant defense by reducing organic hydroperoxides. Sigma-class GSTs metabolize 4-hydroxynonenal which is a by-product of lipid peroxidation produced during the breakdown of long chain lipid hydroperoxides [ 32 , 33 ]. In this context, the observed activity of BaGST2 and BaGST3 toward CuOOH indicates their role in antioxidant defense and metabolism of lipid peroxidation by-product in B. alexandrina .…”

Section: Discussionmentioning

confidence: 99%

Biochemical characterization and peptide mass fingerprinting of two glutathione transferases from Biomphalaria alexandrina snails (Gastropoda: Planorbidae)

Abdalla

Karim

2022

Journal of Genetic Engineering and Biotechnology

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Background The freshwater snails Biomphalaria alexandrina (Gastropoda: Planorbidae) has public health importance of being an intermediate host of Schistosoma mansoni, the parasite species that causes intestinal schistosomiasis in humans. Glutathione transferases (GSTs) play an important role in detoxification of a broad range of compounds including secondary metabolites and exogenous compounds. Studying GSTs in snails may clarify their role in detoxification of molluscicides. Results Two glutathione transferases (BaGST2 and BaGST3) were purified and characterized from B. alexandrina snails. BaGST2 and BaGST3 were electrophoretically homogeneous preparations with subunit molecular weight of 23.6 kDa and molecular weight of 45 kDa. Isoelectric focusing of BaGST2 revealed the presence of two components at pI 4.47 and 4.67, while BaGST3 showed one band at pI 4.17. The specific activity of BaGST2 and BaGST3 toward 1-chloro-2,4-dinitrobenzene (CDNB) was 19.0 and 45.2 μmol/min/mg protein following 146- and 346-fold purification, respectively. The catalytic pH optima, km values, and the activation energies for BaGST2 and BaGST3 were determined. BaGST2 and BaGST3 were significantly inhibited by hematin and Cibacron Blue and to a less extent by bromosulfophthalein, S-butyl-GSH, S-hexyl-GSH, and S-P-bromobenzyl-GSH. BaGST2 and BaGST3 showed high activity against ethacrynic acid as substrate, and they also exhibited peroxidase activity on cumene hydroperoxide. The two enzymes showed identical patterns of lysine-C digestion after high-performance liquid chromatography. The amino acid sequences of three peptide fragments and peptide mass fingerprinting of fourteen peptides were used to predict the primary structure of BaGST2. A polypeptide of 206 amino acids (with 7 gaps, 3 of which could not identified) was predicted for BaGST2. The theoretical subunit molecular weight of BaGST2 is 22.6 kDa, with pI of 8.58. BaGST2 has 65% sequence identity and 78% positive with Biomphalaria glabrata GST7. The overall structure of BaGST2 at the N-terminal domain is identical to the canonical GST N-terminal domain, having the typical thioredoxin-like fold with a βαβ-α-ββα motif, whereas the C-terminal domain is made from 6 α-helices. A conservative GST-N-domain includes glutathione binding sites Y11, L17, Q53, M54, Q65, and S66, while a variable GST-C domain contains electrophilic substrate binding site H99, R102, A103, F106, K107, L161, and Y167. Phylogenetic tree showed that BaGST2 was clustered in the sigma group with GSTs sigma class from invertebrates and vertebrates. Conclusions We have purified and characterized two GSTs from B. alexandrina snails. Our study broadens the biochemical information on freshwater snail GSTs by demonstrating the role of BaGSTs in defense mechanisms against structurally different electrophilic compounds. BaGST2 and BaGST3 have Se-independent peroxidase activity, which indicates their role in cellular antioxidant defense by reducing organic hydroperoxides in B. alexandrina. A polypeptide chain of 206 amino acids was predicted. The primary structure of BaGST2 showed 65% sequence identity with Biomphalaria glabrata GST7. Sequence analysis indicates that BaGST2 is a GST-N-sigma-like with a thioredoxin-like superfamily. Phylogenetic tree confirms that BaGST2 belongs to the sigma class of GSTs superfamily.

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“…A total of 15 Sigma-class GSTs was observed, composing the second-most abundant grouping of GSTs in A. vittatum following the Delta/Epsilon composite group. These enzymes play a role in detoxifying toxic by-products of lipid peroxidation [ 54 , 55 ]. The functional role of Theta-class GSTs is not yet very well understood, although at least in Bombyx mori , these do not appear to play a role in oxidative stress response [ 56 ].…”

Section: Discussionmentioning

confidence: 99%

Transcriptome Sequencing of the Striped Cucumber Beetle, Acalymma vittatum (F.), Reveals Numerous Sex-Specific Transcripts and Xenobiotic Detoxification Genes

Sparks¹,

Nelson

Haber³

et al. 2020

BioTech

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Acalymma vittatum (F.), the striped cucumber beetle, is an important pest of cucurbit crops in the contintental United States, damaging plants through both direct feeding and vectoring of a bacterial wilt pathogen. Besides providing basic biological knowledge, biosequence data for A. vittatum would be useful towards the development of molecular biopesticides to complement existing population control methods. However, no such datasets currently exist. In this study, three biological replicates apiece of male and female adult insects were sequenced and assembled into a set of 630,139 transcripts (of which 232,899 exhibited hits to one or more sequences in NCBI NR). Quantitative analyses identified 2898 genes differentially expressed across the male–female divide, and qualitative analyses characterized the insect’s resistome, comprising the glutathione S-transferase, carboxylesterase, and cytochrome P450 monooxygenase families of xenobiotic detoxification genes. In summary, these data provide useful insights into genes associated with sex differentiation and this beetle’s innate genetic capacity to develop resistance to synthetic pesticides; furthermore, these genes may serve as useful targets for potential use in molecular-based biocontrol technologies.

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Detection and functional characterization of sigma class GST in Phlebotomus argentipes and its role in stress tolerance and DDT resistance

Cited by 27 publications

References 66 publications

Characterization of a sigma class GST (GSTS6) required for cellular detoxification and embryogenesis in Tribolium castaneum

Characterization of a sigma class GST (GSTS6) required for cellular detoxification and embryogenesis in Tribolium castaneum

Biochemical characterization and peptide mass fingerprinting of two glutathione transferases from Biomphalaria alexandrina snails (Gastropoda: Planorbidae)

Transcriptome Sequencing of the Striped Cucumber Beetle, Acalymma vittatum (F.), Reveals Numerous Sex-Specific Transcripts and Xenobiotic Detoxification Genes

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