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Putative ligand binding sites of two functionally characterized bark beetle odorant receptors

Yuvaraj, Jothi K. LU ; Roberts, Rebecca E. LU ; Sonntag, Yonathan LU ; Hou, Xiao-Qing LU ; Grosse-Wilde, Ewald ; Machara, Aleš ; Zhang, Dan-Dan LU ; Hansson, Bill S. ; Johanson, Urban LU orcid and Löfstedt, Christer LU , et al. (2021) In BMC Biology 19.
Abstract

Background: Bark beetles are major pests of conifer forests, and their behavior is primarily mediated via olfaction. Targeting the odorant receptors (ORs) may thus provide avenues towards improved pest control. Such an approach requires information on the function of ORs and their interactions with ligands, which is also essential for understanding the functional evolution of these receptors. Hence, we aimed to identify a high-quality complement of ORs from the destructive spruce bark beetle Ips typographus (Coleoptera, Curculionidae, Scolytinae) and analyze their antennal expression and phylogenetic relationships with ORs from other beetles. Using 68 biologically relevant test compounds, we next aimed to functionally characterize... (More)

Background: Bark beetles are major pests of conifer forests, and their behavior is primarily mediated via olfaction. Targeting the odorant receptors (ORs) may thus provide avenues towards improved pest control. Such an approach requires information on the function of ORs and their interactions with ligands, which is also essential for understanding the functional evolution of these receptors. Hence, we aimed to identify a high-quality complement of ORs from the destructive spruce bark beetle Ips typographus (Coleoptera, Curculionidae, Scolytinae) and analyze their antennal expression and phylogenetic relationships with ORs from other beetles. Using 68 biologically relevant test compounds, we next aimed to functionally characterize ecologically important ORs, using two systems for heterologous expression. Our final aim was to gain insight into the ligand-OR interaction of the functionally characterized ORs, using a combination of computational and experimental methods. Results: We annotated 73 ORs from an antennal transcriptome of I. typographus and report the functional characterization of two ORs (ItypOR46 and ItypOR49), which are responsive to single enantiomers of the common bark beetle pheromone compounds ipsenol and ipsdienol, respectively. Their responses and antennal expression correlate with the specificities, localizations, and/or abundances of olfactory sensory neurons detecting these enantiomers. We use homology modeling and molecular docking to predict their binding sites. Our models reveal a likely binding cleft lined with residues that previously have been shown to affect the responses of insect ORs. Within this cleft, the active ligands are predicted to specifically interact with residues Tyr84 and Thr205 in ItypOR46. The suggested importance of these residues in the activation by ipsenol is experimentally supported through site-directed mutagenesis and functional testing, and hydrogen bonding appears key in pheromone binding. Conclusions: The emerging insight into ligand binding in the two characterized ItypORs has a general importance for our understanding of the molecular and functional evolution of the insect OR gene family. Due to the ecological importance of the characterized receptors and widespread use of ipsenol and ipsdienol in bark beetle chemical communication, these ORs should be evaluated for their potential use in pest control and biosensors to detect bark beetle infestations.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Deorphanization, Functional evolution, HEK293 cells, Odorant receptor, Pest insect, Pheromone receptor, Xenopus oocyte
in
BMC Biology
volume
19
article number
19:16
publisher
BioMed Central (BMC)
external identifiers
  • scopus:85099827932
  • pmid:33499862
ISSN
1741-7007
DOI
10.1186/s12915-020-00946-6
project
The Max Planck Center on next Generation Insect Chemical Ecology
language
English
LU publication?
yes
additional info
Funding Information: This study was funded by the Swedish Research Councils FORMAS (grants #217-2014-689 and #2018-01444 to MNA) and VR (#2017-03804 to CL), the Crafoord foundation (MNA), the Carl Trygger foundation (MNA), the Royal Physiographic Society in Lund (RER and MNA), Stiftelsen Olle Engkvist Byggmästare (YS and UJ), and the Max Planck Society (BSH and EG-W). Open Access funding provided by Lund University. Publisher Copyright: © 2021, The Author(s). Copyright: Copyright 2021 Elsevier B.V., All rights reserved.
id
a1a5df99-912b-4754-b129-99bfcdb8a299
date added to LUP
2021-03-29 08:22:35
date last changed
2025-01-26 07:58:13
@article{a1a5df99-912b-4754-b129-99bfcdb8a299,
  abstract     = {{<p>Background: Bark beetles are major pests of conifer forests, and their behavior is primarily mediated via olfaction. Targeting the odorant receptors (ORs) may thus provide avenues towards improved pest control. Such an approach requires information on the function of ORs and their interactions with ligands, which is also essential for understanding the functional evolution of these receptors. Hence, we aimed to identify a high-quality complement of ORs from the destructive spruce bark beetle Ips typographus (Coleoptera, Curculionidae, Scolytinae) and analyze their antennal expression and phylogenetic relationships with ORs from other beetles. Using 68 biologically relevant test compounds, we next aimed to functionally characterize ecologically important ORs, using two systems for heterologous expression. Our final aim was to gain insight into the ligand-OR interaction of the functionally characterized ORs, using a combination of computational and experimental methods. Results: We annotated 73 ORs from an antennal transcriptome of I. typographus and report the functional characterization of two ORs (ItypOR46 and ItypOR49), which are responsive to single enantiomers of the common bark beetle pheromone compounds ipsenol and ipsdienol, respectively. Their responses and antennal expression correlate with the specificities, localizations, and/or abundances of olfactory sensory neurons detecting these enantiomers. We use homology modeling and molecular docking to predict their binding sites. Our models reveal a likely binding cleft lined with residues that previously have been shown to affect the responses of insect ORs. Within this cleft, the active ligands are predicted to specifically interact with residues Tyr84 and Thr205 in ItypOR46. The suggested importance of these residues in the activation by ipsenol is experimentally supported through site-directed mutagenesis and functional testing, and hydrogen bonding appears key in pheromone binding. Conclusions: The emerging insight into ligand binding in the two characterized ItypORs has a general importance for our understanding of the molecular and functional evolution of the insect OR gene family. Due to the ecological importance of the characterized receptors and widespread use of ipsenol and ipsdienol in bark beetle chemical communication, these ORs should be evaluated for their potential use in pest control and biosensors to detect bark beetle infestations.</p>}},
  author       = {{Yuvaraj, Jothi K. and Roberts, Rebecca E. and Sonntag, Yonathan and Hou, Xiao-Qing and Grosse-Wilde, Ewald and Machara, Aleš and Zhang, Dan-Dan and Hansson, Bill S. and Johanson, Urban and Löfstedt, Christer and Andersson, Martin N.}},
  issn         = {{1741-7007}},
  keywords     = {{Deorphanization; Functional evolution; HEK293 cells; Odorant receptor; Pest insect; Pheromone receptor; Xenopus oocyte}},
  language     = {{eng}},
  month        = {{01}},
  publisher    = {{BioMed Central (BMC)}},
  series       = {{BMC Biology}},
  title        = {{Putative ligand binding sites of two functionally characterized bark beetle odorant receptors}},
  url          = {{http://dx.doi.org/10.1186/s12915-020-00946-6}},
  doi          = {{10.1186/s12915-020-00946-6}},
  volume       = {{19}},
  year         = {{2021}},
}