CYP450s analysis across spiny lobster metamorphosis identifies a long sought missing link in crustacean development

• Published in: The Journal of steroid biochemistry and molecular biology Vol. 171; pp. 262 - 269
• Main Authors: Ventura, Tomer, Bose, Utpal, Fitzgibbon, Quinn P., Smith, Gregory G., Shaw, P. Nicholas, Cummins, Scott F., Elizur, Abigail
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.07.2017; Elsevier BV
• Subjects: Animals; Arthropod Proteins - chemistry; Arthropod Proteins - genetics; Arthropod Proteins - metabolism; Cercopithecus aethiops; Computational Biology; COS Cells; Crustacea; CYP450; Cytochrome; Cytochrome P-450 Enzyme System - chemistry; Cytochrome P-450 Enzyme System - genetics; Cytochrome P-450 Enzyme System - metabolism; Databases, Chemical; Databases, Genetic; Databases, Protein; Decapoda; Ecdysone; Ecdysteroids; Ecdysterone - chemistry; Ecdysterone - metabolism; Enzymes; Gene Expression Profiling; Gene Expression Regulation, Developmental; Genes; Hydroxylation; Juvenile hormone; Metamorphosis; Metamorphosis, Biological; Models, Molecular; Molecular Sequence Annotation; Molecular Structure; Molecular Weight; Molt; Palinuridae - enzymology; Palinuridae - growth & development; Phylogeny; Protein Conformation; Recombinant Proteins - metabolism; Shade; Studies; Ecdysone; Decapoda; CYP450; Metamorphosis; Molt; Juvenile hormone
• ISSN: 0960-0760; 1879-1220
• DOI: 10.1016/j.jsbmb.2017.04.007

•Missing link in crustacean ecdysone hydroxylation identified.•Metamorphic expression implicates un-annotated CYP450s in ecdysteroidogenesis.•CYPome of a spiny lobster enables annotation of known CYP450s based on insects. Cytochrome P450s (CYP450s) are a rapidly evolving family of enzymes, making it difficult to identify bona fide orthologs with notable lineage-specific exceptions. In ecdysozoans, a small number of the most conserved orthologs include enzymes which metabolize ecdysteroids. Ecdysone pathway components were recently shown in a decapod crustacean but with a notable absence of shade, which is important for converting ecdysone to its active form, 20-hydroxyecdysone (20HE), suggesting that another CYP450 performs a similar function in crustaceans. A CYPome temporal expression analysis throughout metamorphosis performed in this research highlights several un-annotated CYP450s displaying differential expression and provides information into expression patterns of annotated CYP450s. Using the expression patterns in the Eastern spiny lobster Sagmariasus verreauxi, followed by 3D modelling and finally activity assays in vitro, we were able to conclude that a group of CYP450s, conserved across decapod crustaceans, function as the insect shade. To emphasize the fact that these genes share the function with shade but are phylogenetically distinct, we name this enzyme system Shed.