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.
Department of Zoology, University of Washington, Seattle 98195.
S R
SR
Palli
K
K
Touhara
J P
JP
Charles
B C
BC
Bonning
J K
JK
Atkinson
S C
SC
Trowell
K
K
Hiruma
W G
WG
Goodman
T
T
Kyriakides
G D
GD
Prestwich
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