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Insect Biochem Mol Biol

Title:		RNA-Seq used to identify ipsdienone reductase (IDONER): A novel monoterpene carbon-carbon double bond reductase central to Ips confusus pheromone production
Author(s):		Fisher KE; Tillett RL; Fotoohi M; Caldwell C; Petereit J; Schlauch K; Tittiger C; Blomquist GJ; MacLean M;
Address:		"Phigenics Research and Innovation Laboratory, Nevada Center for Applied Research, 1664 N. Virginia St., Reno, NV, 89557, USA. Electronic address: kfisher@phigenics.com. Nevada Institute of Personalized Medicine, University of Nevada, Las Vegas, NV, 89154, USA. Electronic address: richard.tillett@unlv.edu. Department of Biochemistry and Molecular Biology, University of Nevada, 1664 N. Virginia St., Reno, NV, 89557, USA. Electronic address: mishafotoohi@yahoo.com. Department of Biochemistry and Molecular Biology, University of Nevada, 1664 N. Virginia St., Reno, NV, 89557, USA. Electronic address: codyacaldwell@yahoo.com. Nevada Center for Bioinformatics, University of Nevada, Reno, NV, 89557, USA. Electronic address: jpetereit@unr.edu. Desert Research Institute, Northern Nevada Science Center Campus, 2215 Raggio Parkway, Reno, NV, 89512, USA. Electronic address: Karen.Schlauch@dri.edu. Department of Biochemistry and Molecular Biology, University of Nevada, 1664 N. Virginia St., Reno, NV, 89557, USA. Electronic address: crt@unr.edu. Department of Biochemistry and Molecular Biology, University of Nevada, 1664 N. Virginia St., Reno, NV, 89557, USA. Electronic address: garyb@cabnr.unr.edu. Department of Biochemistry and Molecular Biology, University of Nevada, 1664 N. Virginia St., Reno, NV, 89557, USA. Electronic address: mmaclean@unr.edu"
Journal Title:		Insect Biochem Mol Biol
Year:		2021
Volume:		20210101
Issue:
Page Number:		103513 -
DOI:		10.1016/j.ibmb.2020.103513
ISSN/ISBN:		1879-0240 (Electronic) 0965-1748 (Print) 0965-1748 (Linking)
Abstract:		"The pinyon ips beetle, Ips confusus (LeConte) is a highly destructive pest in pine forests in western North America. When colonizing a new host tree, I. confusus beetles coordinate a mass attack to overcome the tree's defenses using aggregation pheromones. Ips confusus, as with other Ips spp. beetles, biosynthesize ipsdienol and ipsenol in a specific enantiomeric blend and ratio as aggregation pheromones. While several of the initial steps in the pheromone biosynthetic pathway have been well defined, the final steps were unknown. We used comparative RNA-Seq analysis between fed and unfed male I. confusus midgut tissue to identify candidate genes involved in pheromone biosynthesis. The 12,995 potentially unique transcripts showed a clear separation based on feeding state. Differential expression analysis identified gene groups that were tightly connected. This analysis identified all known pheromone biosynthetic genes and suggested a novel monoterpene double bond reductase, ipsdienone reductase (IDONER), with pheromone biosynthetic gene expression patterns. IDONER cDNA was cloned, expressed, and functionally characterized. The coding DNA sequence has an ORF of 1101 nt with a predicted translation product of 336 amino acids. The enzyme has a molecular weight of 36.7 kDa with conserved motifs of the medium chain dehydrogenases/reductase (MDR) superfamily in the leukotriene B4 dehydrogenases/reductases (LTB(4)R) family. Tagged recombinant protein was expressed and purified. Enzyme assays and GC/MS analysis showed IDONER catalyzed the reduction of ipsdienone to form ipsenone. This study shows that IDONER is a monoterpene double bond reductase involved in I. confusus pheromone biosynthesis"
Keywords:		"Animals Coleoptera/*enzymology Male Monoterpenes/*metabolism Oxidoreductases/*metabolism Pheromones/*biosynthesis Real-Time Polymerase Chain Reaction Sequence Analysis, RNA *Transcriptome Bark beetle Ips confusus Ipsdienone Ipsenone Monoterpene carbon-car;"
Notes:		"MedlineFisher, Katherine E Tillett, Richard L Fotoohi, Misha Caldwell, Cody Petereit, Juli Schlauch, Karen Tittiger, Claus Blomquist, Gary J MacLean, Marina eng P20 GM103440/GM/NIGMS NIH HHS/ P41 GM103311/GM/NIGMS NIH HHS/ U54 GM104944/GM/NIGMS NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Validation Study England 2021/01/04 Insect Biochem Mol Biol. 2021 Feb; 129:103513. doi: 10.1016/j.ibmb.2020.103513. Epub 2021 Jan 1"