Developmental expression of Manduca shade, the P450 mediating the final step in molting hormone synthesis.

Kim Rewitz, Robert Rybczynski, James T. Warren, Lawrence I. Gilbert

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    The ecdysone 20-monooxygenase (E20MO; 20-hydroxylase) is the enzyme that mediates the conversion of ecdysone (E) to the active insect molting hormone, 20-hydroxyecdysone (20E), which coordinates developmental progression. We report the identification and developmental expression of the Halloween gene shade (shd; CYP314A1) that encodes the E20MO in the tobacco hornworm, Manduca sexta. Manduca Shd (MsShd) mediates the conversion of E to 20E when expressed in Drosophila S2 cells. In accord with the central dogma, the data show that Msshd is expressed mainly in the midgut, Malpighian tubules, fat body and epidermis with very low expression in the prothoracic gland and nervous system. Developmental variations in E20MO enzymatic activity are almost perfectly correlated with comparable changes in the gene expression of Msshd in the fat body and midgut during the fifth instar and the beginning of pupal-adult development. The results indicate three successive and overlapping peaks of expression in the fat body, midgut and Malpighian tubules, respectively, during the fifth larval instar. The data suggest that precise tissue-specific transcriptional regulation controls the levels, and thereby the activity, of the Manduca E20MO.
    Original languageEnglish
    JournalMolecular and Cellular Endocrinology
    Volume247
    Issue number1-2
    Pages (from-to)166-174
    Number of pages8
    ISSN0303-7207
    Publication statusPublished - 2006

    Keywords

    • CYP314A1
    • 20-hydroxyecdysone
    • Molting
    • Development
    • Insect
    • Steroid hydroxylation

    Cite this

    Rewitz, Kim ; Rybczynski, Robert ; Warren, James T. ; Gilbert, Lawrence I. / Developmental expression of Manduca shade, the P450 mediating the final step in molting hormone synthesis. In: Molecular and Cellular Endocrinology. 2006 ; Vol. 247, No. 1-2. pp. 166-174.
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    abstract = "The ecdysone 20-monooxygenase (E20MO; 20-hydroxylase) is the enzyme that mediates the conversion of ecdysone (E) to the active insect molting hormone, 20-hydroxyecdysone (20E), which coordinates developmental progression. We report the identification and developmental expression of the Halloween gene shade (shd; CYP314A1) that encodes the E20MO in the tobacco hornworm, Manduca sexta. Manduca Shd (MsShd) mediates the conversion of E to 20E when expressed in Drosophila S2 cells. In accord with the central dogma, the data show that Msshd is expressed mainly in the midgut, Malpighian tubules, fat body and epidermis with very low expression in the prothoracic gland and nervous system. Developmental variations in E20MO enzymatic activity are almost perfectly correlated with comparable changes in the gene expression of Msshd in the fat body and midgut during the fifth instar and the beginning of pupal-adult development. The results indicate three successive and overlapping peaks of expression in the fat body, midgut and Malpighian tubules, respectively, during the fifth larval instar. The data suggest that precise tissue-specific transcriptional regulation controls the levels, and thereby the activity, of the Manduca E20MO.",
    keywords = "CYP314A1, 20-hydroxyecdysone, Molting, Development, Insect, Steroid hydroxylation",
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    Developmental expression of Manduca shade, the P450 mediating the final step in molting hormone synthesis. / Rewitz, Kim; Rybczynski, Robert; Warren, James T.; Gilbert, Lawrence I.

    In: Molecular and Cellular Endocrinology, Vol. 247, No. 1-2, 2006, p. 166-174.

    Research output: Contribution to journalJournal articleResearchpeer-review

    TY - JOUR

    T1 - Developmental expression of Manduca shade, the P450 mediating the final step in molting hormone synthesis.

    AU - Rewitz, Kim

    AU - Rybczynski, Robert

    AU - Warren, James T.

    AU - Gilbert, Lawrence I.

    PY - 2006

    Y1 - 2006

    N2 - The ecdysone 20-monooxygenase (E20MO; 20-hydroxylase) is the enzyme that mediates the conversion of ecdysone (E) to the active insect molting hormone, 20-hydroxyecdysone (20E), which coordinates developmental progression. We report the identification and developmental expression of the Halloween gene shade (shd; CYP314A1) that encodes the E20MO in the tobacco hornworm, Manduca sexta. Manduca Shd (MsShd) mediates the conversion of E to 20E when expressed in Drosophila S2 cells. In accord with the central dogma, the data show that Msshd is expressed mainly in the midgut, Malpighian tubules, fat body and epidermis with very low expression in the prothoracic gland and nervous system. Developmental variations in E20MO enzymatic activity are almost perfectly correlated with comparable changes in the gene expression of Msshd in the fat body and midgut during the fifth instar and the beginning of pupal-adult development. The results indicate three successive and overlapping peaks of expression in the fat body, midgut and Malpighian tubules, respectively, during the fifth larval instar. The data suggest that precise tissue-specific transcriptional regulation controls the levels, and thereby the activity, of the Manduca E20MO.

    AB - The ecdysone 20-monooxygenase (E20MO; 20-hydroxylase) is the enzyme that mediates the conversion of ecdysone (E) to the active insect molting hormone, 20-hydroxyecdysone (20E), which coordinates developmental progression. We report the identification and developmental expression of the Halloween gene shade (shd; CYP314A1) that encodes the E20MO in the tobacco hornworm, Manduca sexta. Manduca Shd (MsShd) mediates the conversion of E to 20E when expressed in Drosophila S2 cells. In accord with the central dogma, the data show that Msshd is expressed mainly in the midgut, Malpighian tubules, fat body and epidermis with very low expression in the prothoracic gland and nervous system. Developmental variations in E20MO enzymatic activity are almost perfectly correlated with comparable changes in the gene expression of Msshd in the fat body and midgut during the fifth instar and the beginning of pupal-adult development. The results indicate three successive and overlapping peaks of expression in the fat body, midgut and Malpighian tubules, respectively, during the fifth larval instar. The data suggest that precise tissue-specific transcriptional regulation controls the levels, and thereby the activity, of the Manduca E20MO.

    KW - CYP314A1

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    KW - Development

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    KW - Steroid hydroxylation

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