Cloning and Sequence Analysis of Rabbit Progesterone-Receptor Complementary DNA

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 83; no. 23; pp. 9045 - 9049
• Main Authors: Loosfelt, Hugues, Atger, Michel, Misrahi, Micheline, Guiochon-Mantel, Anne, Meriel, Cécile, Logeat, Frédérique, Benarous, Richard, Milgrom, Edwin
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.12.1986; National Acad Sciences
• Subjects: Amino Acid Sequence; Amino acids; Animals; Antibodies; Base Sequence; Biological and medical sciences; Biotechnology; Cloning, Molecular; Complementary DNA; Cytosol; DNA; DNA - genetics; Female; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Immunoglobulins; Immunologic Techniques; Messenger RNA; Methods. Procedures. Technologies; Molecular cloning; nucleotide sequence; Oncogene Proteins, Viral; Open reading frames; progesterone; Proteins; Rabbits; Receptors; Receptors, Estrogen - genetics; Receptors, Glucocorticoid - genetics; Receptors, Progesterone - genetics; Receptors, Progesterone - immunology; Recombinant Fusion Proteins - immunology; uterus; Uterus - physiology; vagina; Vertebrata; Mammalia; Complementary DNA; Animal; Rabbit; Primary structure; Progesterone; Lagomorpha; Molecular cloning; Sex steroid hormone; Hormonal receptor
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.83.23.9045

Two λ gt11 clones containing fragments of cDNA encoding the rabbit progesterone receptor were isolated with the aid of monoclonal and monospecific polyclonal antireceptor antibodies. RNA gel blot analysis showed that the corresponding mRNA was ≈ 5900 nucleotides in size and present in the uterus, where its concentration was increased by estrogen treatment, and in the vagina. This mRNA was not detected in liver, in spleen, in intestine, and in kidney where the receptor protein is known to be absent or present in very small concentration. Cross-hybridizing clones were isolated from a λ gt10 library. The DNA was sequenced, and the primary structure of the progesterone receptor was deduced. It consists of 930 amino acids and contains a basic, cysteine-rich region (residues 568-645) with extensive homology to the glucocorticoid and estrogen receptors and the v-erbA oncogene protein. This region is followed by a C-terminal domain that is similar in size to the corresponding domains of the other steroid receptors and v-erbA and shows striking amino acid homology with the glucocorticoid receptor and significant homology with the estrogen receptor. In contrast, the region extending from the cysteine-rich segment toward the N terminus differed in size and amino acid sequence from that of the other receptors and v-erbA. This region had a high proline content in the progesterone receptor.