Molecular cloning and expression of Tenebrio molitor ultraspiracle during metamorphosis and in vivo induction of its phosphorylation by 20-hydroxyecdysone.
Using a RT-PCR approach, the Tenebrio molitor homologue of Drosophila Ultraspiracle (TmUSP) was characterized. Its DNA binding domain shows a degree of identity with those of the other insect USPs. However, the ligand binding domain is closer to those of retinoid X receptors. Using an antibody raised against DmUSP, Western blot analysis of proteins from epidermis and other tissues revealed five immunoreactive bands, corresponding to different phosphorylated forms of a unique polypeptide, as shown by lambda-phosphatase treatment. The nuclear form of TmUSP seems unphosphorylated. An in vivo 20-hydroxyecdysone treatment increases considerably and rapidly the phosphorylated forms of TmUSP. This post-translational modification may play a role in the 20-hydroxyecdysone response.
Nicolaï
M
M
UMR CNRS 5548 Développement-Communication Chimique, Université de Bourgogne, Dijon, France.
Bouhin
H
H
Quennedey
B
B
Delachambre
J
J
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins; D004441:Ecdysterone / Q000494:pharmacology*; D004817:Epidermis / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D008675:Metamorphosis, Biological; D008969:Molecular Sequence Data; D010766:Phosphorylation; D020033:Protein Isoforms / Q000145:classification / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger; D017422:Sequence Analysis, DNA / Q000379:methods; D013712:Tenebrio / Q000235:genetics* / Q000254:growth & development; D014157:Transcription Factors / Q000145:classification / Q000235:genetics* / Q000378:metabolism
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger; D014157:Transcription Factors; C066030:USP protein, Drosophila; D004441:Ecdysterone