{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T02:59:20Z","timestamp":1787540360665,"version":"build-2736575974"},"reference-count":54,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2022,7,16]],"date-time":"2022-07-16T00:00:00Z","timestamp":1657929600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Juvenile hormone epoxide hydrolase (JHEH) plays an important role in the metabolism of JH III in insects. To study the control of JHEH in female Drosophila melanogaster, JHEH 1, 2 and 3 cDNAs were cloned and sequenced. Northern blot analyses showed that the three transcripts are expressed in the head thorax, the gut, the ovaries and the fat body of females. Molecular modeling shows that the enzyme is a homodimer that binds juvenile hormone III acid (JH IIIA) at the catalytic groove better than JH III. Analyses of the three JHEH promoters and expressing short promoter sequences behind a reporter gene (lacZ) in D. melanogaster cell culture identified a JHEH 3 promoter sequence (626 bp) that is 10- and 25-fold more active than the most active promoter sequences of JHEH 2 and JHEH 1, respectively. A transcription factor (TF) Sp1 that is involved in the activation of JHEH 3 promoter sequence was identified. Knocking down Sp1 using dsRNA inhibited the transcriptional activity of this promoter in transfected D. melanogaster cells and JH III and 20HE downregulated the JHEH 3 promoter. On the other hand, JH IIIA and farnesoic acid did not affect the promoter, indicating that JH IIIA is JHEH\u2019s preferred substrate. A transgenic D. melanogaster expressing a highly activated JHEH 3 promoter behind a lacZ reporter gene showed promoter transcriptional activity in many D. melanogaster tissues.<\/jats:p>","DOI":"10.3390\/biom12070991","type":"journal-article","created":{"date-parts":[[2022,7,17]],"date-time":"2022-07-17T21:00:28Z","timestamp":1658091628000},"page":"991","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Cloning and Characterization of Drosophila melanogaster Juvenile Hormone Epoxide Hydrolases (JHEH) and Their Promoters"],"prefix":"10.3390","volume":"12","author":[{"given":"Dov","family":"Borovsky","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hilde","family":"Breyssens","sequence":"additional","affiliation":[{"name":"Zoological Institute, KU Leuven, 3000 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Esther","family":"Buytaert","sequence":"additional","affiliation":[{"name":"Zoological Institute, KU Leuven, 3000 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tom","family":"Peeters","sequence":"additional","affiliation":[{"name":"Zoological Institute, KU Leuven, 3000 Leuven, Belgium"},{"name":"Open BioLab Brussels, Erasmushogeschool Brussels, 1210 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carole","family":"Laroye","sequence":"additional","affiliation":[{"name":"Zoological Institute, KU Leuven, 3000 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karolien","family":"Stoffels","sequence":"additional","affiliation":[{"name":"Zoological Institute, KU Leuven, 3000 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9936-7843","authenticated-orcid":false,"given":"Pierre","family":"Roug\u00e9","sequence":"additional","affiliation":[{"name":"Faculte des Sciences Pharmaceutiques, 31400 Tolouse, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/S0065-2806(08)60084-3","article-title":"Cellular and molecular actions of juvenile hormone general considerations and premetamorphic actions","volume":"24","author":"Riddiford","year":"1994","journal-title":"Adv. Insect Physiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0065-2806(08)60030-2","article-title":"Cellular and molecular actions of juvenile hormone. II. Roles of juvenile hormone in adult insects","volume":"26","author":"Wyatt","year":"1996","journal-title":"Adv. Insect Physiol."},{"key":"ref_3","first-page":"267","article-title":"Allatostatins: Identification, primary structure, function and distribution","volume":"125","author":"Stay","year":"1994","journal-title":"Adv. Insect Physiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1016\/0003-2697(77)90196-8","article-title":"A rapid assay for insect juvenile hormone esterase activity","volume":"82","author":"Hammock","year":"1977","journal-title":"Anal. Biochem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/0003-2697(88)90257-6","article-title":"A partition assay for the simultaneous determination of insect juvenile hormone esterase and epoxide hydrolase activity","volume":"169","author":"Share","year":"1988","journal-title":"Anal. Biochem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"619","DOI":"10.1093\/jmedent\/29.4.619","article-title":"In vivo and in vitro biosynthesis and metabolism of methyl farnesoate, juvenile hormone III, and juvenile hormone III acid in the mosquito Aedes aegypti","volume":"29","author":"Borovsky","year":"1992","journal-title":"J. Med. Entomol."},{"key":"ref_7","first-page":"431","article-title":"Guidelines for the use and interpretation of assays for monitoring autophagy","volume":"Volume 7","author":"Kerkut","year":"1985","journal-title":"Comprehensive Insect Physiology, Biochemistry and Pharmacology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"11751","DOI":"10.1073\/pnas.0500954102","article-title":"Precocious metamorphosis in transgenic silkworms overexpressing juvenile hormone esterase","volume":"102","author":"Tan","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.jinsphys.2014.03.001","article-title":"Aedes aegypti juvenile hormone acid methyl transferase, the ultimate enzyme in the biosynthetic pathway of juvenile hormone III, exhibits substrate control","volume":"64","author":"Heylen","year":"2014","journal-title":"J. Insect Physiol."},{"key":"ref_10","unstructured":"Gupta, A.P. (1990). Metabolism of juvenile hormones: Degradation and titer regulation. Morphogenetic Hormones of Arthropods, Rutgers University Press."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5234","DOI":"10.1021\/bi00015a037","article-title":"Key disulfide bonds in an insect hormone binding protein: cDNA cloning of a juvenile hormone binding protein of Heliothis virescens and ligand binding by native and mutant forms","volume":"34","author":"Wojtasek","year":"1995","journal-title":"Biochemistry"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1016\/j.jinsphys.2004.12.008","article-title":"Regulation of JH epoxide hydrolase versus JH esterase activity in the cabbage looper, Trichoplusia ni, by juvenile hormone and xenobiotics","volume":"51","author":"Anspaugh","year":"2005","journal-title":"J. Insect Physiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1016\/0965-1748(92)90045-G","article-title":"Characterization of juvenile hormone esterase in Drosophila melanogaster","volume":"22","author":"Campbell","year":"1992","journal-title":"Insect Biochem. Mol. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"988","DOI":"10.1007\/BF02125646","article-title":"Juvenile hormone catabolism in Manduca sexta: Homologue selectivity of catabolism and identification of a diol-phosphate conjugate as a major end product","volume":"49","author":"Halarnkar","year":"1993","journal-title":"Experientia"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1016\/0022-1910(95)00037-U","article-title":"Juvenile hormone metabolism during the fourth stadium and pupal stage of the Southern House Mosquito Culex quinquefasciatus","volume":"41","author":"Lassiter","year":"1995","journal-title":"J. Insect Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3224","DOI":"10.1002\/prot.24676","article-title":"Crystal structure of juvenile hormone epoxide hydrolase from the silkworm Bombyx mori","volume":"82","author":"Zhou","year":"2014","journal-title":"Proteins Struct. Funct. Bioinform."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1006\/bbrc.1996.0404","article-title":"An insect juvenile hormone-specific epoxide hydrolase is related to vertebrate microsomal epoxide hydrolases","volume":"220","author":"Wojtasek","year":"1996","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1016\/S0965-1748(01)00141-2","article-title":"cDNA cloning and characterization of Bombyx mori juvenile hormone esterase: An inducible gene by the imidazole insect growth regulator KK-42","volume":"32","author":"Hirai","year":"2002","journal-title":"Insect Biochem. Mol. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1139","DOI":"10.1016\/j.jinsphys.2010.03.007","article-title":"The juvenile hormone (JH) epoxide hydrolase gene in the honeybee (Apis mellifera) genome encodes a protein which has negligible participation in JH degradation","volume":"56","author":"Mackert","year":"2010","journal-title":"J. Insect Physiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4696","DOI":"10.1046\/j.1432-1033.2003.03868.x","article-title":"Characterization and cDNA cloning of a clofibrate-inducible microsomal epoxide hydrolase in Drosophila melanogaster","volume":"270","author":"Taniai","year":"2003","journal-title":"Eur. J. Biochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1146\/annurev.ento.43.1.671","article-title":"Differential gene expression in insects: Transcriptional control","volume":"43","author":"Harshman","year":"1998","journal-title":"Annu. Rev. Entomol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1146\/annurev-ento-010814-020810","article-title":"Chorion genes: A landscape of their evolution, structure, and regulation","volume":"60","author":"Papantonis","year":"2015","journal-title":"Annu. Rev. Entomol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1573","DOI":"10.1016\/j.cell.2017.11.008","article-title":"YY1 is a structural regulator of enhancer-promoter loops","volume":"171","author":"Weintraub","year":"2017","journal-title":"Cell"},{"key":"ref_24","first-page":"7004","article-title":"Identifcation of a novel strong promoter from the anhydrobiotic midge, Polypedilum vanderplanki, with conserved function in various insect cell lines","volume":"9","author":"Miyata","year":"2019","journal-title":"Nat. Sci. Rep."},{"key":"ref_25","first-page":"2363","article-title":"Identification and functional analysis of promoters of heat-shock genes from the fall armyworm, Spodoptera frugiperda","volume":"10","author":"Chen","year":"2020","journal-title":"Nat. Sci. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Bleckmann, M., Fritz, M.H.-Y., Bhuju, S., Jarek, M., Sch\u00fcrig, M., Geffers, R., Benes, V., Besir, H., and van den Heuvel, J. (2015). Genomic Analysis and Isolation of RNA Polymerase II Dependent Promoters from Spodoptera frugiperda. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0132898"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1006\/bbrc.1993.2508","article-title":"Insect immunity: The diptericin promoter contains multiple functional regulatory sequences homologous to mammalian acute-phase response elements","volume":"197","author":"Georgel","year":"1993","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1140","DOI":"10.1093\/nar\/23.7.1140","article-title":"Drosophila immunity. A sequence homologous to mammalian interferon consensus response element enhances the activity of the diptericin promoter","volume":"23","author":"Georgel","year":"1995","journal-title":"Nucleic Acids Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e9816","DOI":"10.15252\/msb.20209816","article-title":"Large-scale analysis of Drosophila core promoter function using synthetic promoters","volume":"18","author":"Qi","year":"2022","journal-title":"Mol. Syst. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1016\/0378-1119(88)90177-1","article-title":"Vectors for Drosophila P-element-mediated transformation and tissue culture transformation","volume":"74","author":"Thummel","year":"1988","journal-title":"Gene"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1093\/jmedent\/29.2.318","article-title":"In vitro assay for biosynthesis and metabolism of juvenile hormone by exposed corpora allata of Aedes aegypti (diptera: Culicidae)","volume":"20","author":"Borovsky","year":"1992","journal-title":"J. Med. Entomol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1002\/(SICI)1520-6327(1996)32:3\/4<527::AID-ARCH24>3.0.CO;2-D","article-title":"Mechanism of Action and Cloning of Epoxide Hydrolase From the Cabbage Looper, Trichoplusia ni","volume":"32","author":"Roe","year":"1996","journal-title":"Arch. Insect Biochem. Physiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1016\/S0965-1748(98)00014-9","article-title":"Expression and characterization of the recombinant juvenile hormone epoxide hydrolase (JHEH) from Manduca sexta","volume":"28","author":"Debernard","year":"1998","journal-title":"Insect Biochem. Mol. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1016\/0076-6879(93)18026-9","article-title":"Rapid amplification of complementary DNA ends for generation of full length complementary DNAs: Thermal RACE","volume":"8","author":"Frohman","year":"1993","journal-title":"Methods Enzymol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1111\/j.1432-1033.1996.0279n.x","article-title":"Molecular sequencing and modeling of Neobellieria bullata trypsin: Evidence for translational control with Neb TMOF","volume":"237","author":"Borovsky","year":"1996","journal-title":"Eur. J. Biochem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1046\/j.1432-1327.1999.00411.x","article-title":"Sequencing and characterization of the citrus weevil Diaprepes abbreviatus, trypsin cDNA; effect of Aedes trypsin modulating oostatic factor on trypsin biosynthesis","volume":"262","author":"Yan","year":"1999","journal-title":"Eur. J. Biochem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1002\/prot.10104","article-title":"Increasing the precision of comparative models with YASARA NOVA-a self-parameterizing force field","volume":"57","author":"Krieger","year":"2002","journal-title":"Proteins"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"7334","DOI":"10.1021\/ja809673d","article-title":"Directed evolution of an enantioselective epoxide hydrolase: Uncovering the source of enantioselectivity at each evolutionary stage","volume":"131","author":"Reetz","year":"2009","journal-title":"J. Am. Chem. Soc."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/S0969-2126(00)00087-3","article-title":"Structure of Aspergillus niger epoxide hydrolase at 1.8 A resolution: Implications for the structure and function of the mammalian microsomal class of epoxide hydrolases","volume":"8","author":"Zou","year":"2000","journal-title":"Structure"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1107\/S0021889892009944","article-title":"Procheck: A program to check the stereochemistry of protein structures","volume":"26","author":"Laskowski","year":"1993","journal-title":"J. Appl. Cryst."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1006\/jmbi.1998.1665","article-title":"Assessing protein structures with a non-local atomic interaction energy","volume":"277","author":"Melo","year":"1998","journal-title":"J. Mol. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1093\/bioinformatics\/btq662","article-title":"Toward the estimation of the absolute quality of individual protein structure models","volume":"27","author":"Benkert","year":"2011","journal-title":"Bioinformatics"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1093\/bioinformatics\/bti770","article-title":"The SWISS-MODEL workspace: A web-based environment for protein structure homology modelling","volume":"22","author":"Arnold","year":"2006","journal-title":"Bioinformatics"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2149","DOI":"10.1002\/jcc.21797","article-title":"Fast docking using the CHARMM force field with EADock DSS","volume":"32","author":"Grosdidier","year":"2011","journal-title":"J. Comput. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1605","DOI":"10.1002\/jcc.20084","article-title":"UCSF Chimera-a visualization system for exploratory research and analysis","volume":"25","author":"Pettersen","year":"2004","journal-title":"J. Comput. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/S0097-8485(01)00099-7","article-title":"Application of a time-delay neural network to promoter annotation in the Drosophila melanogaster genome","volume":"26","author":"Reese","year":"2001","journal-title":"Comput. Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1093\/jisesa\/iey002","article-title":"Characterization and Expression Profiles of Juvenile Hormone Epoxide Hydrolase From Lymantria dispar (Lepidoptera: Lymantridae) and RNA Interference by Ingestion","volume":"18","author":"Wen","year":"2018","journal-title":"J. Insect Sci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1038\/s41598-017-00907-0","article-title":"Juvenile Hormone Epoxide Hydrolase: A Promising Target for Hemipteran Pest Management","volume":"7","author":"Tusun","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1016\/0020-1790(91)90089-W","article-title":"cDNA and deduced amino acid sequence of a blood meal-induced trypsin from the mosquito Aedes aegypti","volume":"21","author":"Graf","year":"1991","journal-title":"Insect Biochem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1073\/pnas.1716897115","article-title":"Antagonistic actions of juvenile hormone and 20-hydroxyecdysone within the ring gland determine developmental transitions in Drosophila","volume":"115","author":"Liu","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"180239","DOI":"10.1098\/rsob.180239","article-title":"Post-translational modification control of RNA-binding protein hnRNPK function","volume":"9","author":"Xu","year":"2019","journal-title":"Open Biol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1111\/j.1432-1033.1987.tb11184.x","article-title":"The titre of juvenile hormone during the pupal and adult stages of the life cycle of Drosophila melanogaster","volume":"164","author":"Bownes","year":"1987","journal-title":"Eur. J. Biochem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0083-6729(00)60016-X","article-title":"Ecdysone receptors and their biological actions","volume":"60","author":"Riddiford","year":"2000","journal-title":"Vitam. Horm."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/j.jinsphys.2004.12.007","article-title":"Developmental and hormonal regulation of juvenile hormone esterase gene in Drosophila melanogaster","volume":"51","author":"Kethidi","year":"2005","journal-title":"J. Insect Physiol."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/12\/7\/991\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:51:52Z","timestamp":1760140312000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/12\/7\/991"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,16]]},"references-count":54,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["biom12070991"],"URL":"https:\/\/doi.org\/10.3390\/biom12070991","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,16]]}}}