A conserved hymenopteran-specific family of cytochrome P450s protects bee pollinators from toxic nectar alkaloids

Haas, Julian; Beck, Elena; Troczka, Bartlomiej J.; Hayward, Angela; Hertlein, Gillian; Zaworra, Marion; Lueke, Bettina; Buer, Benjamin; Maiwald, Frank; Beck, Michael Edmund; Nebelsiek, Birgit; Glaubitz, Johannes; Bass, Chris; Nauen, Ralf

doi:10.1126/sciadv.adg0885

Cited by 10 publications

(3 citation statements)

References 91 publications

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“…For example, cytochrome P450s of the CYP6B gene subfamily endowed the genus Papilio (swallowtail butterflies) with the ability to feed on furanocoumarin-containing plants ( 43 ). Similarly, P450s of the 336A family were recently identified as a conserved mechanism that allowed honeybees and other hymenopteran species that diverged over 281 Mya to metabolize alkaloids ( 44 ). The loss of the key DIMBOA-metabolizing UGT in S. picta , and its resulting high sensitivity to this PSM, is consistent with its specialization on Crinum plants which do not produce this PSM.…”

Section: Discussionmentioning

confidence: 99%

UDP-glycosyltransferases act as key determinants of host plant range in generalist and specialist Spodoptera species

Wang,

Song,

Hunt

et al. 2024

Proc. Natl. Acad. Sci. U.S.A.

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Phytophagous insects have evolved sophisticated detoxification systems to overcome the antiherbivore chemical defenses produced by many plants. However, how these biotransformation systems differ in generalist and specialist insect species and their role in determining insect host plant range remains an open question. Here, we show that UDP-glucosyltransferases (UGTs) play a key role in determining the host range of insect species within the Spodoptera genus. Comparative genomic analyses of Spodoptera species that differ in host plant breadth identified a relatively conserved number of UGT genes in generalist species but high levels of UGT gene pseudogenization in the specialist Spodoptera picta . CRISPR-Cas9 knockouts of the three main UGT gene clusters of Spodoptera frugiperda revealed that UGT33 genes play an important role in allowing this species to utilize the poaceous plants maize, wheat, and rice, while UGT40 genes facilitate utilization of cotton. Further functional analyses in vivo and in vitro identified the UGT SfUGT33F32 as the key mechanism that allows generalist S. frugiperda to detoxify the benzoxazinoid DIMBOA (2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one), a potent insecticidal phytotoxin produced by poaceous plants. However, while this detoxification capacity is conserved in several generalist Spodoptera species, Spodoptera picta , which specializes on Crinum plants, is unable to detoxify DIMBOA due to a nonfunctionalizing mutation in SpUGT33F34 . Collectively, these findings provide insight into the role of insect UGTs in host plant adaptation, the mechanistic basis of evolutionary transitions between generalism and specialism and offer molecular targets for controlling a group of notorious insect pests.

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Section: Discussionmentioning

confidence: 99%

UDP-glycosyltransferases act as key determinants of host plant range in generalist and specialist Spodoptera species

Wang,

Song,

Hunt

et al. 2024

Proc. Natl. Acad. Sci. U.S.A.

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“…Work on human CYP3A4, has demonstrated the importance of key amino acid residues in the SRSs, including E 308 from the SRS4/M2 motif (Kiani et al., 2019 ). Further, a single charged residue in SRS4 (D 298 ), in the CYP336A P450 family across the Hymenoptera, has also been shown to act as a gatekeeper that facilitates the specificity of the enzyme (Haas et al., 2023 ). As such, the loss of the anionic residue in the CYP9DM enzymes (equivalent to E 308 in CYP3A4), provides an indication that these enzymes bind a different group of substrates to those of the CYP9Q‐related lineage.…”

Section: Discussionmentioning

confidence: 99%

A cytochrome P450 insecticide detoxification mechanism is not conserved across the Megachilidae family of bees

Hayward,

Hunt,

Haas

et al. 2023

Evolutionary Applications

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Recent work has demonstrated that many bee species have specific cytochrome P450 enzymes (P450s) that can efficiently detoxify certain insecticides. The presence of these P450s, belonging or closely related to the CYP9Q subfamily (CYP9Q‐related), is generally well conserved across the diversity of bees. However, the alfalfa leafcutter bee, Megachile rotundata, lacks CYP9Q‐related P450s and is 170–2500 times more sensitive to certain insecticides than bee pollinators with these P450s. The extent to which these findings apply to other Megachilidae bee species remains uncertain. To address this knowledge gap, we sequenced the transcriptomes of four Megachile species and leveraged the data obtained, in combination with publicly available genomic data, to investigate the evolution and function of P450s in the Megachilidae. Our analyses reveal that several Megachilidae species, belonging to the Lithurgini, Megachilini and Anthidini tribes, including all species of the Megachile genus investigated, lack CYP9Q‐related genes. In place of these genes Megachile species have evolved phylogenetically distinct CYP9 genes, the CYP9DM lineage. Functional expression of these P450s from M. rotundata reveal they lack the capacity to metabolize the neonicotinoid insecticides thiacloprid and imidacloprid. In contrast, species from the Osmiini and Dioxyini tribes of Megachilidae have CYP9Q‐related P450s belonging to the CYP9BU subfamily that are able to detoxify thiacloprid. These findings provide new insight into the evolution of P450s that act as key determinants of insecticide sensitivity in bees and have important applied implications for pesticide risk assessment.

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“…The natural bond orbital (NBO) spin population was obtained at the UB3LYP/B2 level. The Grimme DFT-D3BJ corrections were added to all QM/MM calculations. − As both our calculations (refer to Tables S2 and S3) and previous studies show that S = 1/2 (doublet) and S = 3/2 (quartet) states exhibit generally similar reactivities for the Cpd I-mediated reactions, ,− our discussions are limited to the doublet state in the main text. The calculated spin populations of key atoms of the involved species are shown in Table S1.…”

Section: Methodsmentioning

confidence: 99%

Substrate Conformational Switch Enables the Stereoselective Dimerization in P450 NascB: Insights from Molecular Dynamics Simulations and Quantum Mechanical/Molecular Mechanical Calculations

Zhou,

Feng,

Wang

et al. 2024

JACS Au

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P450 NascB catalyzes the coupling of cyclo-(l-tryptophan-l-proline) (1) to generate (−)-naseseazine C (2) through intramolecular C–N bond formation and intermolecular C–C coupling. A thorough understanding of its catalytic mechanism is crucial for the engineering or design of P450-catalyzed C–N dimerization reactions. By employing MD simulations, QM/MM calculations, and enhanced sampling, we assessed various mechanisms from recent works. Our study demonstrates that the most favorable pathway entails the transfer of a hydrogen atom from N7–H to Cpd I. Subsequently, there is a conformational change in the substrate radical, shifting it from the Re-face to the Si-face of N7 in Substrate 1. The Si-face conformation of Substrate 1 is stabilized by the protein environment and the π–π stacking interaction between the indole ring and heme porphyrin. The subsequent intermolecular C3–C6′ bond formation between Substrate 1 radical and Substrate 2 occurs via a radical attack mechanism. The conformational switch of the Substrate 1 radical not only lowers the barrier of the intermolecular C3–C6′ bond formation but also yields the correct stereoselectivity observed in experiments. In addition, we evaluated the reactivity of the ferric-superoxide species, showing it is not reactive enough to initiate the hydrogen atom abstraction from the indole NH group of the substrate. Our simulation provides a comprehensive mechanistic insight into how the P450 enzyme precisely controls both the intramolecular C–N cyclization and intermolecular C–C coupling. The current findings align with the available experimental data, emphasizing the pivotal role of substrate dynamics in governing P450 catalysis.

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A conserved hymenopteran-specific family of cytochrome P450s protects bee pollinators from toxic nectar alkaloids

Cited by 10 publications

References 91 publications

UDP-glycosyltransferases act as key determinants of host plant range in generalist and specialist Spodoptera species

UDP-glycosyltransferases act as key determinants of host plant range in generalist and specialist Spodoptera species

A cytochrome P450 insecticide detoxification mechanism is not conserved across the Megachilidae family of bees

Substrate Conformational Switch Enables the Stereoselective Dimerization in P450 NascB: Insights from Molecular Dynamics Simulations and Quantum Mechanical/Molecular Mechanical Calculations

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