Open AccessIsolation of mRNA from Bovine Pituitary
Author(s) -
KELLER David,
FETHERSTON Jackie,
BOIME Irving
Publication year - 1980
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1980.tb04731.x
Subject(s) - immunoprecipitation , protein biosynthesis , alpha (finance) , beta (programming language) , messenger rna , g alpha subunit , microbiology and biotechnology , biology , protein subunit , rna , biochemistry , luteinizing hormone , gel electrophoresis , antiserum , methionine , translation (biology) , hormone , amino acid , gene , antibody , medicine , construct validity , nursing , patient satisfaction , programming language , computer science , immunology
RNA derived from bovine steer pituitary was translated in wheat germ cell‐free extracts containing [ 35 S]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.