Molecular evolution of the insect Halloween family of cytochrome P450s: Phylogeny, gene organization and functional conservation

Kim F. Rewitz, Michael B. O'Connor, Lawrence I. Gilbert

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178 Scopus citations


The insect molting hormone, 20-hydroxyecdysone (20E), is a major modulator of the developmental processes resulting in molting and metamorphosis. During evolution selective forces have preserved the Halloween genes encoding cytochrome P450 (P450) enzymes that mediate the biosynthesis of 20E. In the present study, we examine the phylogenetic relationships of these P450 genes in holometabolous insects belonging to the orders Hymenoptera, Coleoptera, Lepidoptera and Diptera. The analyzed insect genomes each contains single orthologs of Phantom (CYP306A1), Disembodied (CYP302A1), Shadow (CYP315A1) and Shade (CYP314A1), the terminal hydroxylases. In Drosophila melanogaster, the Halloween gene spook (Cyp307a1) is required for the biosynthesis of 20E, although a function has not yet been identified. Unlike the other Halloween genes, the ancestor of this gene evolved into three paralogs, all in the CYP307 family, through gene duplication. The genomic stability of these paralogs varies among species. Intron-exon structures indicate that D. melanogaster Cyp307a1 is a mRNA-derived paralog of spookier (Cyp307a2), which is the ancestral gene and the closest ortholog of the coleopteran, lepidopteran and mosquito CYP307A subfamily genes. Evolutionary links between the insect Halloween genes and vertebrate steroidogenic P450s suggest that they originated from common ancestors, perhaps destined for steroidogenesis, before the deuterostome-arthropod split. Conservation of putative substrate recognition sites of orthologous Halloween genes indicates selective constraint on these residues to prevent functional divergence. The results suggest that duplications of ancestral P450 genes that acquired novel functions may have been an important mechanism for evolving the ecdysteroidogenic pathway.

Original languageEnglish (US)
Pages (from-to)741-753
Number of pages13
JournalInsect Biochemistry and Molecular Biology
Issue number8 SPEC. ISS
StatePublished - Aug 2007

Bibliographical note

Funding Information:
We thank members of the Gilbert and O’Connor laboratories for their consistent and excellent work on the Halloween gene project over the past several years as well as their collegiality. In addition we are indebted to Todd Vision (Bioinformaticist, University of North Carolina) and Rene Feyereisen (INRA) for helpful comments on the manuscript and Joseph Shaw (Dartmouth and Mt. Desert Island Biological Laboratory) for his help in gaining entry to the Daphnia database. We are also thankful to Ole Andersen and Anders Løbner-Olesen (Roskilde University). This work was supported by Grant IBN130825 from the National Science Foundation to LIG. MBO is an investigator of the Howard Hughes Medical Institute.


  • 20-Hydroxyecdysone
  • Ecdysteroids
  • Gene duplication
  • Molting
  • P450
  • Steroid hydroxylation


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