title: Molecular cloning of the prothoracicotropic hormone from the tobacco hornworm, Manduca sexta.
abstract: A cDNA encoding a putative precursor of prothoracicotropic hormone (PTTH) from the tobacco hornworm, Manduca sexta, was isolated and sequenced. This clone contains an open reading frame encoding a 226-amino acid prepropeptide hormone. The deduced amino acid sequence is composed of a signal sequence, a precursor domain and a mature hormone and shows similarities to the other PTTHs that have been cloned from closely related lepidopteran species, Bombyx mori, Samia cynthia ricini, Antheraea peryni, and Hyalophora cecropia. Although these cDNAs showed slightly less similarities in predicted amino acid sequences, seven cysteine residues and the hydrophobic regions within those mature peptides were conserved. In situ hybridization using a cDNA probe encoding the Manduca PTTH showed that PTTH mRNA was in two pairs of neurosecretory cells in the Manduca brain. The recombinant putative Manduca PTTH produced in E. coli was biologically active, both causing a larval molt in neck-ligated Manduca 4th instar larvae (ED(50)=50 pM) and the adult molt of diapausing Manduca pupae (ED(50)=79 pM), but was unable to stimulate molting of debrained Bombyx pupae.
authors.affiliation: Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Kashiwanoha 5-1-5, Kashiwa 277-8562, Japan.
authors.forename: Motoko
authors.initials: M
authors.lastname: Shionoya
authors.forename: Hideki
authors.initials: H
authors.lastname: Matsubayashi
authors.forename: Masako
authors.initials: M
authors.lastname: Asahina
authors.forename: Hisato
authors.initials: H
authors.lastname: Kuniyoshi
authors.forename: Shinji
authors.initials: S
authors.lastname: Nagata
authors.forename: Lynn M
authors.initials: LM
authors.lastname: Riddiford
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kataoka
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006023:Glycoproteins; D017403:In Situ Hybridization; D007301:Insect Hormones / Q000235:genetics*; D007814:Larva / Q000254:growth & development / Q000502:physiology; D018402:Manduca / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D018978:Molting / Q000502:physiology; D009479:Neuropeptides; D017422:Sequence Analysis, DNA
chemical_list: D018076:DNA, Complementary; D006023:Glycoproteins; D007301:Insect Hormones; D009479:Neuropeptides; C009089:prothoracicotropic hormone