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. 1979 Jan;76(1):298–302. doi: 10.1073/pnas.76.1.298

mRNA-directed biosynthesis of α subunit of thyrotropin: Translation in cell-free and whole-cell systems

Ione A Kourides *, Bruce D Weintraub †
PMCID: PMC382926  PMID: 284345

Abstract

mRNA from mouse thyrotropic pituitary tumors was translated in frog oocytes (a whole-cell system) and in wheat germ extract and reticulocyte lysate (cell-free systems) in the presence of [35S]methionine. Synthesized peptides related to thyrotropin were identified in the three systems by immunoprecipitation with subunit-specific antisera developed against the α subunit of ovine lutropin (luteinizing hormone) and the β subunit of bovine thyrotropin. In wheat germ extract and reticulocyte lysate, a single immunoprecipitable form of the α subunit of thyrotropin was synthesized with an apparent molecular weight of 14,000 by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. In the frog oocyte, three forms of immunoprecipitable α subunit of thyrotropin were synthesized with apparent molecular weights of 20,000, 14,000, and 10,000. The 20,000 form is similar to unlabeled rat pituitary standard α subunit and 35S-labeled mouse tumor α subunit in cell cultures (20,000-21,000); thus, it may represent a precursor-cleaved and glycosylated form. The 14,000 form synthesized in all three systems probably represents the pre-α subunit of thyrotropin; the 10,000 form, synthesized only in the frog oocyte, could be a proteolytically cleaved but unglycosylated form. Because only the α subunit of thyrotropin was identified and no larger molecular weight immunoprecipitable form of either subunit was detected in any of the translation systems, α and β subunits of thyrotropin appear to be translated from separate mRNAs.

Keywords: secretory glycoprotein hormone, mouse thyrotropic pituitary tumor, independent synthesis of thyrotropin subunits

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Selected References

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