Characterization of crab EcR and RXR homologs and expression during limb regeneration and oocyte maturation.
We report here complete coding sequences for the Uca pugilator homologs of the ecdysteroid (UpEcR) and retinoid-X receptors (UpRXR). Library screenings recovered cDNA clones containing a unique amino terminal open-reading frame (A/B domain) for each gene, most similar to insect B1 EcR and USP1/RXR isoforms. Splicing variants in the UpRXR ligand-binding domain were also identified, in a region critical for folding of Drosophila and lepidopteran USP. UpEcR and UpRXR proteins were able to associate, and both are required for binding to an ecdysteroid HRE; these interactions were not hormone-dependent. Ribonuclease protection assays (RPA) were conducted using A/B domain and 'common' (C or E) domain probes on RNA isolated from various stages of regenerating limb buds and ovaries. For several of the limb bud and ovarian stages examined, the relative level of A/B domain sequence protected was significantly less than common domain suggesting alternative amino terminal isoforms other than those isolated through cloning. This is the first report of UpEcR and UpRXR transcription during ovarian maturation, implicating the ovary as a potential target for hormonal control in Crustacea.
Durica
David S
DS
Department of Zoology, University of Oklahoma, 730 Van Vleet Oval, Norman, OK 73019, USA. ddurica@ou.edu
Wu
Xiaohui
X
Anilkumar
Gopinathan
G
Hopkins
Penny M
PM
Chung
Arthur C-K
AC
D000595:Amino Acid Sequence; D000818:Animals; D003386:Brachyura / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005121:Extremities / Q000502:physiology*; D015723:Gene Library; D017930:Genes, Reporter; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism; D007447:Invertebrate Hormones / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D009865:Oocytes / Q000502:physiology*; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism*; D011987:Receptors, Steroid / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012038:Regeneration / Q000502:physiology*; D047488:Retinoid X Receptors; D016415:Sequence Alignment; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; D019476:Insect Proteins; D007447:Invertebrate Hormones; D009687:Nuclear Proteins; D018168:Receptors, Retinoic Acid; D011987:Receptors, Steroid; D011993:Recombinant Fusion Proteins; D047488:Retinoid X Receptors; D014157:Transcription Factors; C034808:ecdysteroid receptor