Control of juvenile hormone biosynthesis in Bombyx mori:: Cloning of the enzymes in the mevalonate pathway and assessment of their developmental expression in the corpora allata

We have isolated the cDNAs of all enzymes involved in the mevalonate pathway portion of the juvenile hormone (JH) biosynthetic pathway in Bombyx mori, i.e., those responsible for the formation of farnesyl diphosphate from acetyl-CoA. There is a single gene encoding each enzyme of this pathway, with the exception of farnesyl diphosohate synthase (FPPS), for which we identified three hornologs. All but two of these enzymes are expressed almost exclusively in the corpora allata (CA), as indicated by quantitative RT-PCR analyses. Phosphornevalonate kinase (MevPK) was expressed in many tissues, including the CA. In day 24th instars, FPPS1 expression was detected primarily in the Malpighian tubules, but expression of the structurally related FPPS2 and FPPS3 occurred mainly in the CA. Since FPPS3 transcripts were 55 times less abundant than those of FPPS2, the latter is expecied to play a major role in JH biosynthesis at this stage. Studies on the developmental expression of these enzymes in the CA showed that the levels of all transcripts were high during the 4th instar larvae, a stage at which in vitro JH biosynthesis was high. However, the transcripts of all the mevalonate enzymes declined to low levels and JH acid O-methyltransferase (JHAMT) transcript disappeared by day 3 when CA ceased JH production after the final larval molt. The CA did not synthesize JH during the pupal stage, coincident with the limited expression of mevalonate kinase, phosphornevalonate kinase, diphosphornevalonate kinase and isopentenyl diphosphate isomerase, and the inactivation of the JHANIT gene. Only female CA produced JH in the adult stage, a feature associated with the re-expression of JHAMT in female but little in male adult CA. Altogether, our results point to a relationship between JH biosynthesis and expression of most JH biosynthetic enzymes in the CA. (c) 2007 Elsevier Ltd. All rights reserved.