Isolation of mRNA from Bovine Pituitary: The Cell‐Free Synthesis of the α and β Subunits of Luteinizing Hormone

David KELLER, Jackie FETHERSTON, Irving BOIME

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Abstract

RNA derived from bovine steer pituitary was translated in wheat germ cell‐free extracts containing [35S]methionine. Antisera generated against purified denatured α and β subunits of lutropin were used to demonstrate the synthesis of both proteins in vitro. The immunoprecipitated products of the cell‐free system were resolved on sodium dodecyl sulfate/polyacrylamide gels and it was observed that the molecular weight of the immunoprecipitated α subunit protein was approximately 14000, while that of the β protein was estimated to be 16000. Since the molecular weights of authentic α and β subunits are 10600 and 14000 respectively, the cell‐free products presumably represented their pre‐protein forms. The ratio of the immunoprecipitated subunit pre‐proteins was dependent on the magnesium concentration in the translation mixtures; at 2.1 mM, translation of lutropin α and β mRNAs was comparable. RNA isolated from cow pituitary tissue directed the synthesis of fivefold less of the α and β immunoprecipitated proteins than did steer RNA. Since the blood levels of gonadal steroids are higher in the cow, the results supported the hypothesis that lutropin α and β mRNA biosynthesis is repressed by these steroids. The data also suggest that synthesis of lutropin α and β subunits is coordinately expressed in certain physiological situations.

Original languageEnglish
Pages (from-to)367-372
Number of pages6
JournalEuropean Journal of Biochemistry
Volume108
Issue number2
DOIs
StatePublished - Jul 1980

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