Two functionally distinct CYP4G genes of Anopheles gambiae contribute to cuticular hydrocarbon biosynthesis

Kefi, Mary; Balabanidou, Vasileia; Douris, Vassilis; Lycett, Gareth; Vontas, John

doi:10.1016/j.ibmb.2019.04.018

Cited by 50 publications

(42 citation statements)

References 34 publications

(67 reference statements)

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“…Most recently, knockdown of CYP4G14 in Tribolium castaneum caused the beetles to be more susceptible to tefluthrin . Among these genes, only CYP4G16 and CYP4G17 have been functionally characterized as the oxidative decarbonylase for hydrocarbon biosynthesis, and have been suggested to play a role in penetration resistance in A. gambiae . The association of other CYP4G members with hydrocarbon biosynthesis and their roles in insecticide resistance require future investigation.…”

Section: Discussionmentioning

confidence: 99%

“…58 Among these genes, only CYP4G16 and CYP4G17 have been functionally characterized as the oxidative decarbonylase for hydrocarbon biosynthesis, and have been suggested to play a role in penetration resistance in A. gambiae. 9,32 The association of other CYP4G members with hydrocarbon biosynthesis and their roles in insecticide resistance require future investigation. The current work, together with the studies of CYP4Gs on A. gambiae, prompts speculation that CYP4G-mediated penetration resistance via the enrichment of CHCs might be a universal resistance mechanism across contact insecticide resistant insects.…”

Section: Discussionmentioning

confidence: 99%

“…Cytochromes P450 are remarkable for their diverse functions, and their roles in insecticide resistance are mostly associated with metabolic resistance by enhancing detoxification . However, two CYP4G subfamily genes, CYP4G16 and CYP4G17 , have been reported to be positively associated with cuticular resistance via catalyzing the production of CHCs in A. gambiae . The only CYP4G member in B. germanica , CYP4G19 , was reported to be overexpressed in a pyrethroid‐resistant strain, but no role in insecticide resistance has been attributed to CYP4G19 .…”

Section: Introductionmentioning

confidence: 99%

“…31 However, two CYP4G subfamily genes, CYP4G16 and CYP4G17, have been reported to be positively associated with cuticular resistance via catalyzing the production of CHCs in A. gambiae. 9,32 The only CYP4G member in B. germanica, CYP4G19, 33 was reported to be overexpressed in a pyrethroid-resistant strain, 34 but no role in insecticide resistance has been attributed to CYP4G19. Several CYP4G genes have been functionally characterized to be involved in the final decarbonylation step of hydrocarbon biosynthesis by baculovirus expression system 9,[35][36][37] or RNAi-based functional studies.…”

Section: Introductionmentioning

confidence: 99%

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Involvement of integument‐rich CYP4G19 in hydrocarbon biosynthesis and cuticular penetration resistance in Blattella germanica (L.)

Chen

Pei²,

et al. 2019

Pest Management Science

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BACKGROUND Cuticle penetration plays an important role as a mechanism of insecticide resistance, but the underlying molecular mechanism remains poorly understood. In Blattella germanica, the cytochrome P450 gene, CYP4G19, is overexpressed in a pyrethroid‐resistant strain. Here, we investigated whether CYP4G19 is involved in the biosynthesis of hydrocarbons and further contributes to cuticular penetration resistance in B. germanica. RESULTS Compared with the susceptible strain, pyrethroid‐resistant cockroaches showed lower cuticular permeability with Eosin Y staining. Removal of epicuticular lipids, mainly nonpolar hydrocarbons, with a hexane wash intensified the cuticular permeability and decreased the resistance index of the resistant strain. CYP4G19 was predominately expressed in the abdominal integument and could be upregulated by desiccation stress or short exposure to beta‐cypermethrin. Overexpression of CYP4G19 in the resistant strain was positively correlated with a higher level of cuticular hydrocarbons (CHCs). RNAi‐mediated knockdown of CYP4G19 significantly decreased its expression and caused a reduction in CHCs. Meanwhile, CYP4G19 suppression resulted in a non‐uniform array of the lipid layer, enhanced cuticle permeability, and compromised insecticide tolerance. CONCLUSION Our findings confirm that CYP4G19 is involved in hydrocarbon production and appears to contribute to hydrocarbon‐based penetration resistance in B. germanica. This study highlights the lipid‐based penetration resistance, advancing our understanding of the molecular mechanism underlying P450‐mediated cuticular penetration resistance in insects. © 2019 Society of Chemical Industry

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Involvement of integument‐rich CYP4G19 in hydrocarbon biosynthesis and cuticular penetration resistance in Blattella germanica (L.)

Chen

Pei²,

et al. 2019

Pest Management Science

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“…During later pupal to adult development, CYP4G17 appears dominate in the production of methyl branched chains. A propensity to generate methyl-CHCs was also observed when cyp4g17 is ectopically expressed in cyp4G1 silenced in Drosophila (Kefi et al, 2019). It should be noted however that the temporal analysis is complicated by the unknown rate of deposition of internally stored CHCs to the cuticle.…”

Section: Discussionmentioning

confidence: 92%

Development of a functional genetic tool for Anopheles gambiae oenocyte characterisation: application to cuticular hydrocarbon synthesis

Lynd

Balabanidou

Grosman

et al. 2019

Preprint

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27Oenocytes are an insect cell type having diverse physiological functions ranging 28 from cuticular hydrocarbon (CHC) production to insecticide detoxification that may 29 impact their capacity to transmit pathogens. To develop functional genetic tools to 30 study Anopheles gambiae oenocytes, we have trapped an oenocyte enhancer to 31 create a transgenic mosquito Gal4 driver line that mediates tissue-specific 32 expression. After crossing with UAS-reporter lines, An. gambiae oenocytes are 33 fluorescently tagged through all life stages and demonstrate clearly the two 34 characteristic oenocyte cell-types arising during development. The driver was then 35 used to characterise the function of two oenocyte expressed An. gambiae cyp4g 36 genes through tissue-specific expression of UAS-RNAi constructs. Silencing of 37 cyp4g16 or cyp4g17 caused lethality in pupae of differing timing and penetrance. 38 Surviving cyp4g16 knockdown adults showed increased sensitivity to desiccation. 39 Total cuticular hydrocarbon levels were reduced by approximately 80% or 50% in 40 both single gene knockdowns when assayed in young pupa or surviving adults 41 respectively, indicating both genes are required for complete CHC production in An. 42 gambiae and demonstrate synergistic activity in young pupae. Comparative CHC 43 profiles were very similar for the two knockdowns, indicating overlapping substrate 44 specificities of the two enzymes. Differences were observed for example with 45 reduced abundance of shorter chain CHCs in CYP4G16 knockdowns, and reduction 46 in longer, branched chained CHCs in CYP4G17 knockdown adults. This is the first 47 time that two cyp4gs have both been shown to be required for complete CHC 48 production in an insect. Moreover, the generation of tagged cells and identification of 49 an enhancer region can expediate oenocyte specific transcriptomics. The novel 50 4 driver line can also be used to explore oenocyte roles in pheromone production, 51 mating behaviour and longevity in the malaria mosquito. 52 53 54

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Both LmCYP4G genes function in decreasing cuticular penetration of insecticides in Locusta migratoria

Zhang²,

et al. 2020

Pest Management Science

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BACKGROUND: Cuticular hydrocarbons (CHCs) have a critical role in preventing desiccation and penetration of xenobiotics in insects. Previous studies have shown that cytochrome P450 subfamily 4G (CYP4G) enzymes are oxidative decarbonylases, essential for CHC biosynthesis. However, it is unclear whether there are functional differences between the two CYP4G genes in most insects. In Locusta migratoria, we identified two CYP4G genes (LmCYP4G62 and LmCYP4G102). LmCYP4G102 plays a critical role in the synthesis of CHCs, but the function of LmCYP4G62 is unknown. RESULTS: We identified, characterized, and compared two LmCYP4G genes, based on L. migratoria transcriptomic and genomic databases. RT-qPCR showed that both were highly expressed in tissues with which oenocytes are associated, the integument and fat body. Immunostaining indicated that LmCYP4G62 and LmCYP4G102 were highly abundant in oenocytes in these tissues. However, the two enzymes had a different subcellular distribution, with LmCYP4G62 localized on the plasma membrane and LmCYP4G102 dispersed throughout the oenocyte cytoplasm, presumably on the endoplasmic reticulum. RNA interferencemediated gene silencing against each of the two genes resulted in reduced CHC contents, in all classes for LmCYP4G102, but mostly shorter chain CHCs for LmCYP4G62. Silencing of both genes resulted in increased insecticide penetration through the cuticle, and increased locust susceptibility to desiccation and insecticides. CONCLUSION: Our studies suggest that both LmCYP4G62 and LmCYP4G102 contribute to hydrocarbon biosynthesis and play key roles in protecting locusts from water loss and insecticide penetration, but they are not fully redundant. Further, the two LmCYP4G genes might be used as new targets for insect pest management.

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Two functionally distinct CYP4G genes of Anopheles gambiae contribute to cuticular hydrocarbon biosynthesis

Cited by 50 publications

References 34 publications

Involvement of integument‐rich CYP4G19 in hydrocarbon biosynthesis and cuticular penetration resistance in Blattella germanica (L.)

Involvement of integument‐rich CYP4G19 in hydrocarbon biosynthesis and cuticular penetration resistance in Blattella germanica (L.)

Development of a functional genetic tool for Anopheles gambiae oenocyte characterisation: application to cuticular hydrocarbon synthesis

Both LmCYP4G genes function in decreasing cuticular penetration of insecticides in Locusta migratoria

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