Cell-free translation of the messenger RNA for calf terminal deoxynucleotidyltransferase

• Published in: The Journal of biological chemistry 1982-04, Vol.257 (8), p.4013-4015
• Main Authors: Wolf, S C, Kourides, I A, Good, R A, Silverstone, A E
• Format: Article
• Language: English
• Subjects: Animals; Antigen-Antibody Complex; Cattle; DNA Nucleotidylexotransferase - genetics; DNA Nucleotidylexotransferase - isolation & purification; DNA Nucleotidyltransferases - genetics; Immune Sera; Molecular Weight; Poly A - genetics; Protein Biosynthesis; Rabbits; Reticulocytes - metabolism; RNA - genetics; RNA, Messenger - genetics; Thymus Gland - enzymology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)34674-X

Poly(A)-rich RNA has been isolated from calf thymus and translated in vitro in a rabbit reticulocyte translation system. Three peptides with Mr = 58,000, 33,000, and 13,000 were specifically immunoprecipitated from the translation products with calf terminal deoxynucleotidyltransferase antiserum. An oligo(dT)-purified preparation of calf terminal transferase competed with only the Mr = 58,000 peptide in the immunoprecipitation reaction. The anti-terminal transferase serum did not precipitate a Mr = 58,000 peptide from translation products of spleen or liver mRNA, but it did precipitate the Mr = 33,000 and 13,000 peptides from products of spleen mRNA and a Mr = 13,000 peptide from products of liver mRNA. In addition, when an affinity-purified antibody to calf terminal transferase was used, only a Mr = 58,000 peptide was immunoprecipitated from the translation products of calf thymus mRNA, and none was immunoprecipitated from spleen or liver mRNA products. This antibody also precipitated a Mr = 58,000 peptide from the cell lysates of calf thymocytes labeled in vitro with [35S]methionine. These results demonstrate that calf terminal transferase is biosynthesized as a Mr = 58,000 peptide.