Early steps in ecdysteroid biosynthesis: Evidence for the involvement of cytochrome P-450 enzymes

Grieneisen, Michael L.; Warren, James T.; Gilbert, Lawrence I.

doi:10.1016/0965-1748(93)90077-6

Cited by 96 publications

(53 citation statements)

References 43 publications

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“…Previous biochemical studies have shown that the conversion of C to 7dC is mediated by an enzyme with cytochrome P450 characteristics under the control of the Zn-finger protein Without Children (Woc) (Grieneisen et al, 1993;Warren et al, 1995;Warren et al, 2001). Our RT-PCR analysis did not detect a significant difference of nvd-Dm expression level between woc mutant and wild type (see Fig.…”

Section: Molecular Functions Of the Neverland Family Of Proteinsmentioning

confidence: 46%

“…We performed the feeding experiment with two precursors of ecdysteroid biosynthesis, cholesterol (C) and 7-dehydrocholesterol (7dC) . C and 7dC are involved in the first step of ecdysteroidogenesis in the PG; 7dC is synthesized from C by 7,8-dehydrogenation by an uncharacterized enzyme (Grieneisen et al, 1993;Gilbert et al, 2002;Gilbert and Warren, 2005). When 7dC was added to the food, all nvd-Dm RNAi animals were completely rescued and developed into adults (Fig.…”

Section: The Lethality Of Nvd-dm Rnai Animals Is Completely Rescued Bmentioning

confidence: 99%

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Neverland is an evolutionally conserved Rieske-domain protein that is essential for ecdysone synthesis and insect growth

et al. 2006

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Steroid hormones mediate a wide variety of developmental and physiological events in multicellular organisms. During larval and pupal stages of insects, the principal steroid hormone is ecdysone, which is synthesized in the prothoracic gland (PG) and plays a central role in the control of development. Although many studies have revealed the biochemical features of ecdysone synthesis in the PG, many aspects of this pathway have remained unclear at the molecular level. We describe the neverland (nvd) gene, which encodes an oxygenase-like protein with a Rieske electron carrier domain, from the silkworm Bombyx mori and the fruitfly Drosophila melanogaster. nvd is expressed specifically in tissues that synthesize ecdysone, such as the PG. We also show that loss of nvd function in the PG causes arrest of both molting and growth during Drosophila development. Furthermore, the phenotype is rescued by application of 20-hydroxyecdysone or the precursor 7-dehydrocholesterol. Given that the nvd family is evolutionally conserved, these results suggest that Nvd is an essential regulator of cholesterol metabolism or trafficking in steroid synthesis across animal phyla.

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Section: Molecular Functions Of the Neverland Family Of Proteinsmentioning

confidence: 46%

Section: The Lethality Of Nvd-dm Rnai Animals Is Completely Rescued Bmentioning

confidence: 99%

Neverland is an evolutionally conserved Rieske-domain protein that is essential for ecdysone synthesis and insect growth

et al. 2006

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“…The latter two tissues were also used for subsequent in vitro incubation with various radiolabeled intermediates (8,9). Prothoracic glands were dissected from late fifth instar Manduca sexta larvae and frozen (Ϫ70°C) until thawed and incubated with various radiolabeled intermediates (10)(11)(12) resulting sequence, both mutant sad alleles were identified by the appearance of two bases at a given position in the sequence.…”

Section: Methodsmentioning

confidence: 99%

“…Equipment and procedures for RP-HPLC and normal phase TLC have been described previously (8)(9)(10)(11)(12). For MS analysis, after sequential RP-HPLC and TLC purification of (2).…”

Section: Riamentioning

confidence: 99%

Molecular and biochemical characterization of two P450 enzymes in the ecdysteroidogenic pathway of Drosophila melanogaster

Warren

Petryk

Marqués

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

306

243

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Five different enzymatic activities, catalyzed by both microsomal and mitochondrial cytochrome P450 monooxygenases (CYPs), are strongly implicated in the biosynthesis of ecdysone (E) from cholesterol. However, none of these enzymes have been characterized completely. The present data show that the wild-type genes of two members of the Halloween family of embryonic lethals, disembodied (dib) and shadow (sad), code for mitochondrial cytochromes P450 that mediate the last two hydroxylation reactions in the ecdysteroidogenic pathway in Drosophila, namely the C 22-and C 2-hydroxylases. When sad (CYP315A1) is transfected into Drosophila S2 cells, the cells metabolize 2-deoxyecdysone (2dE) H]E in high yield. The expression of sad and dib is concentrated within the individual segments of the developing epidermis when there is a surge of ecdysteroid midway through embryogenesis. This result occurs before the ring gland has developed and suggests that the embryonic epidermis is a site of ecdysteroid biosynthesis. This pattern then diminishes, and, during late embryogenesis, expression of both genes is concentrated in the prothoracic gland cells of the developing ring gland. Expression of dib and sad continues to be localized in this endocrine compartment during larval development, being maximal in both the late second and third instar larvae, about the time of the premolt peaks in the ecdysteroid titer.

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“…Cytochrome P450s are involved in insect growth and development through the processing of such compounds as pheromones and ecdysteroids (4,5). They have also been implicated in metabolic resistance to insecticides, including carbamates, chlorinated hydrocarbons, organophosphates, and pyrethroids (6,7).…”

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confidence: 99%

Molecular cloning of a family of xenobiotic-inducible drosophilid cytochrome P450s: Evidence for involvement in host-plant allelochemical resistance

Danielson

MacIntyre

Fogleman

1997

Proc. Natl. Acad. Sci. U.S.A.

114

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Cytochrome P450s constitute a superfamily of genes encoding mostly microsomal hemoproteins that play a dominant role in the metabolism of a wide variety of both endogenous and foreign compounds. In insects, xenobiotic metabolism (i.e., metabolism of insecticides and toxic natural plant compounds) is known to involve members of the CYP6 family of cytochrome P450s. Use of a 3 RACE (rapid amplification of cDNA ends) strategy with a degenerate primer based on the conserved cytochrome P450 heme-binding decapeptide loop resulted in the amplification of four cDNA sequences representing another family of cytochrome P450 genes (CYP28) from two species of isoquinoline alkaloidresistant Drosophila and the cosmopolitan species Drosophila hydei. The CYP28 family forms a monophyletic clade with strong regional homologies to the vertebrate CYP3 family and the insect CYP6 family (both of which are involved in xenobiotic metabolism) and to the insect CYP9 family (of unknown function). Induction of mRNA levels for three of the CYP28 cytochrome P450s by toxic host-plant allelochemicals (up to 11.5-fold) and phenobarbital (up to 49-fold) corroborates previous in vitro metabolism studies and suggests a potentially important role for the CYP28 family in determining patterns of insect-host-plant relationships through xenobiotic detoxification.

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Early steps in ecdysteroid biosynthesis: Evidence for the involvement of cytochrome P-450 enzymes

Cited by 96 publications

References 43 publications

Neverland is an evolutionally conserved Rieske-domain protein that is essential for ecdysone synthesis and insect growth

Neverland is an evolutionally conserved Rieske-domain protein that is essential for ecdysone synthesis and insect growth

Molecular and biochemical characterization of two P450 enzymes in the ecdysteroidogenic pathway of Drosophila melanogaster

Molecular cloning of a family of xenobiotic-inducible drosophilid cytochrome P450s: Evidence for involvement in host-plant allelochemical resistance

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