Molecular cloning and characterization of growth hormone receptor and its homologue in the Japanese eel (Anguilla japonica).
Two cDNAs encoding growth hormone receptor (GHR)-like genes, eGHR1 and eGHR2, were isolated from Japanese eel (Anguilla japonica) liver tissue. The putative eel GHR proteins showed conserved structural features of vertebrate GHRs, including six cysteine residues and a YGEFS motif in the extracellular domain, a single transmembrane region, and proline-rich box 1 and box 2 domains. Northern blot analysis showed a single eGHR1 transcript in liver, while two sizes of eGHR2 transcripts, thought to be produced by alternative splicing, were present. RT-PCR revealed that eGHR1 and eGHR2 transcripts were widely distributed throughout the whole body of the Japanese eel. Moreover, the results of binding assays showed the specific binding of growth hormone to recombinant eGHR1. Since these putative eGHR proteins show all characteristics of the GHR family, we conclude that eGHR1 and eGHR2 cDNA encode two different GHRs in Japanese eel. We confirmed the ligand specificity of eGHR1 by binding assay, and further research is needed to allow characterization of the binding capability of eGHR2.
Ozaki
Yuichi
Y
Division of Marine Biosciences, Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido 041-8611, Japan. ozaki-yu@agr.ehime-u.ac.jp
Fukada
Haruhisa
H
Kazeto
Yukinori
Y
Adachi
Shinji
S
Hara
Akihiko
A
Yamauchi
Kohei
K
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D004524:Eels / Q000235:genetics*; D005260:Female; D029941:Fish Proteins / Q000235:genetics*; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D009928:Organ Specificity; D017434:Protein Structure, Tertiary; D011986:Receptors, Somatotropin / Q000235:genetics*
D029941:Fish Proteins; D011986:Receptors, Somatotropin