A nuclear juvenile hormone-binding protein from larvae of Manduca sexta: a putative receptor for the metamorphic action of juvenile hormone.
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 (Kd = 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.
Palli
S R
SR
Department of Zoology, University of Washington, Seattle 98195.
Touhara
K
K
Charles
J P
JP
Bonning
B C
BC
Atkinson
J K
JK
Trowell
S C
SC
Hiruma
K
K
Goodman
W G
WG
Kyriakides
T
T
Prestwich
G D
GD
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D018076:DNA, Complementary / Q000302:isolation & purification; D019476:Insect Proteins; D007536:Isomerism; D007605:Juvenile Hormones / Q000378:metabolism; D007700:Kinetics; D007814:Larva; D008297:Male; D008675:Metamorphosis, Biological / Q000502:physiology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009036:Moths / Q000254:growth & development / Q000378:metabolism* / Q000502:physiology; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods; D012717:Sesquiterpenes / Q000378:metabolism; D014316:Tritium
D002352:Carrier Proteins; D018076:DNA, Complementary; D019476:Insect Proteins; D007605:Juvenile Hormones; D009838:Oligodeoxyribonucleotides; D012717:Sesquiterpenes; C035606:juvenile hormone-binding protein, insect; D014316:Tritium; C008442:juvenile hormone I