A nuclear juvenile hormone-binding protein from larvae of Manduca sexta: A putative receptor for the metamorphic action of juvenile hormone

Subba R. Palli, Kazushige Touhara, Jean Philippe Charles, Bryony C. Bonning, Jeffrey K. Atkinson, Stephen C. Trowell, Kiyoshi Hiruma, Walter G. Goodman, Themis Kyriakides, Glenn D. Prestwich, Bruce D. Hammock, Lynn M. Riddiford

Research output: Contribution to journalArticle

54 Scopus citations

Abstract

A 29-kDa nuclear juvenile hormone (JH)-binding protein from the epidermis of Manduca sexta larvae was purified by using the photoaffinity analog for JH II ([3H]epoxyhomofarnesyldiazoacetate) and partially sequenced. A 1.1-kb cDNA was isolated by using degenerate oligonucleotide primers for PCR based on these sequences. The cDNA encoded a 262-amino acid protein that showed no similarity with other known proteins, except for short stretches of the interphotoreceptor retinoid-binding protein, rhodopsin, and human nuclear protein p68. Recombinant baculovirus containing this cDNA made a 29-kDa protein that was covalently modified by [3H]epoxyhomofarnesyldiazoacetate and specifically bound the natural enantiomer of JH I (K(d) = 10.7 nM). This binding was inhibited by the natural JHs but not by methoprene. Immunocytochemical analysis showed localization of this 29-kDa protein to epidermal nuclei. Both mRNA and protein are present during the intermolt periods; during the larval molt, the mRNA disappears but the protein persists. Later when cells become pupally committed, both the mRNA and protein disappear with a transient reappearance near pupal ecdysis. The properties of this protein are consistent with its being the receptor necessary for the antimetamorphic effects of JH.

Original languageEnglish (US)
Pages (from-to)6191-6195
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume91
Issue number13
DOIs
StatePublished - Jun 21 1994

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Keywords

  • baculovirus expression
  • metamorphosis
  • photoaffinity labeling

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Palli, S. R., Touhara, K., Charles, J. P., Bonning, B. C., Atkinson, J. K., Trowell, S. C., Hiruma, K., Goodman, W. G., Kyriakides, T., Prestwich, G. D., Hammock, B. D., & Riddiford, L. M. (1994). A nuclear juvenile hormone-binding protein from larvae of Manduca sexta: A putative receptor for the metamorphic action of juvenile hormone. Proceedings of the National Academy of Sciences of the United States of America, 91(13), 6191-6195. https://doi.org/10.1073/pnas.91.13.6191