Expression Cloning of a cDNA Encoding a Fish Prolactin Receptor

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 92; no. 13; pp. 6037 - 6041
• Main Authors: Sandra, Olivier, Sohm, Frederic, De Luze, Amaury, Prunet, Patrick, Edery, Marc, Kelly, Paul A.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 20.06.1995; National Acad Sciences; National Academy of Sciences
• Subjects: Amino Acid Sequence; Amino acids; Animals; Base Sequence; Birds; Cattle; Cell Line; Cellular Biology; Cercopithecus aethiops; Chickens; Cloning, Molecular; Columbidae; Complementary DNA; COS cells; Deoxyribonucleic acid; DNA; DNA, Complementary - biosynthesis; DNA, Complementary - metabolism; Fish; Freshwater; Gene Expression; Gene Library; Generally accepted auditing standards; Genetics; Hormones; Humans; Kidney; Kidneys; Kinetics; Life Sciences; Mice; Molecular Sequence Data; Oreochromis niloticus; Prolactin receptors; Rabbits; Rats; Receptors; Receptors, Prolactin - biosynthesis; Receptors, Prolactin - metabolism; Recombinant Proteins - biosynthesis; Recombinant Proteins - metabolism; RNA, Messenger - analysis; RNA, Messenger - biosynthesis; Sequence Homology, Amino Acid; Tilapia; Transfection; Vertebrates; BIOLOGIE MOLECULAIRE
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.92.13.6037

By using an expression cloning strategy, we isolated a single positive clone encoding a tilapia prolactin (PRL) receptor. Tilapia PRL188was used to screen a fresh-water tilapia kidney expression library transfected in COS cells. The tilapia PRL receptor is a mature protein of 606 amino acids. The extracellular domain is devoid of the tandem repeat units present in birds and has two pairs of cysteine residues, a Trp-Ser-Xaa-Trp-Ser motif, and two potential N-glycosylation sites. The cytoplasmic domain contains 372 amino acids, including box 1, a sequence previously shown to be important for signal transduction in mammalian species. Thus, the general structure is similar to the long form of mammalian PRL receptors; however, amino acid comparisons reveal a rather low identity (≈37%). Northern blot analysis shows the existence of a single transcript in osmoregulatory tissues and reproductive organs. This localization is in agreement with known functions of PRL in teleosts.