Cloning and Characterization of cDNAs Encoding Human Gastrin-Releasing Peptide

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 81; no. 18; pp. 5699 - 5703
• Main Authors: Spindel, Eliot R., Chin, William W., Price, Janet, Rees, Lesley H., Besser, Gordon M., Habener, Joel F.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 01.09.1984; National Acad Sciences
• Subjects: Amino Acid Sequence; Amino acids; Base Sequence; Basic amino acids; Biochemistry; Bombesin - genetics; Carcinoid Tumor - genetics; Carcinoid tumors; Cloning, Molecular; Complementary DNA; DNA - metabolism; DNA Restriction Enzymes; Gastrin-Releasing Peptide; Humans; Liver Neoplasms - genetics; Liver Neoplasms - secondary; Lungs; Messenger RNA; Nucleic Acid Hybridization; Peptides - genetics; Plasmids; Poly A - isolation & purification; RNA; RNA - isolation & purification; RNA, Messenger - genetics; Tumors
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.81.18.5699

We have prepared and cloned cDNAs derived from poly(A)+RNA from a human pulmonary carcinoid tumor rich in immunoreactivity to gastrin-releasing peptide, a peptide closely related in structure to amphibian bombesin. Mixtures of synthetic oligodeoxyribonucleotides corresponding to amphibian bombesin were used as hybridization probes to screen a cDNA library prepared from the tumor RNA. Sequencing of the recombinant plasmids shows that human gastrin-releasing peptide (hGRP) mRNA encodes a precursor of 148 amino acids containing a typical signal sequence, hGRP consisting of 27 or 28 amino acids, and a carboxyl-terminal extension peptide. hGRP is flanked at its carboxyl terminus by two basic amino acids, following a glycine used for amidation of the carboxyl-terminal methionine. RNA blot analyses of tumor RNA show a major mRNA of 900 bases and a minor mRNA of 850 bases. Blot hybridization analyses using human genomic DNA are consistent with a single hGRP-encoding gene. The presence of two mRNAs encoding the hGRP precursor protein in the face of a single hGRP gene raises the possibility of alternative processing of the single RNA transcript.