Juvenile hormone regulation of Drosophila Epac--a guanine nucleotide exchange factor.
Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to characterize the effects of juvenile hormone (JH) on Epac (Exchange Protein directly Activated by Cyclic AMP; NM_001103732), a guanine nucleotide exchange factor for Rap1 in Drosophila S2 cells. JH treatment led to a rapid, dose-dependent increase in Epac relative expression ratio (RER) when compared to treatment with methyl linoleate (MLA) that lacks biological activity. The minimal level of hormone needed to elicit a response was 100 ng/ml. Time-course studies indicated a significant rise in the RER 1h after treatment. S2 cells were challenged with 20-hydroxyecdysone and a series of compounds similar in structure to JH to determine the specificity of the response. Methoprene and JH III displayed the greatest increases in RER. Late third instar (96 h) Drosophila were exposed to diet containing methoprene (500 ng/g diet); significantly higher RERs for Epac were observed 12h after exposure. JH had no effect on Epac RERs in the human cell line HEK-293.
Wang
Jun
J
Department of Entomology, University of Wisconsin-Madison, Madison, WI 53706, United States.
Lindholm
Joliene R
JR
Willis
David K
DK
Orth
Anthony
A
Goodman
Walter G
WG
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004330:Drosophila / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004441:Ecdysterone / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007605:Juvenile Hormones / Q000494:pharmacology*; D007814:Larva / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D008726:Methoprene / Q000494:pharmacology; D012717:Sesquiterpenes / Q000494:pharmacology; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism
D029721:Drosophila Proteins; C542725:Epac protein, Drosophila; D020662:Guanine Nucleotide Exchange Factors; D007605:Juvenile Hormones; D012717:Sesquiterpenes; D004441:Ecdysterone; D008726:Methoprene; C036585:juvenile hormone III; D020827:rap1 GTP-Binding Proteins