METHYLMERCURY HYDROXIDE ENHANCEMENT OF TRANSLATION AND TRANSCRIPTION OF OVALBUMIN AND CONALBUMIN MESSENGER-RNAS

Abstract

Translation of total mRNA in heterologous protein-synthesizing systems is often employed as an indirect means of assessing relative mRNA concentrations. The efficiency of translation of specific mRNA differs. One such example is the poor translational efficiency of conalbumin mRNA relative to [hen oviduct] ovalbumin mRNA. The translation of conalbumin and ovalbumin mRNA in crude mRNA preparations and with highly purified mRNA preparations was studied. Treatment of RNA with methylmercury hydroxide prior to translation improves the translational efficiency of both mRNA and preferentially improves translational efficiency of conalbumin mRNA to the point where it more correctly reflects the relative concentration of these 2 mRNA in crude mRNA preparations. Conalbumin mRNA is also a poor template for the synthesis of full length cDNA synthesis by avian myeloblastosis virus reverse transcriptase, and treatment of this mRNA with methylmercury hydroxide increases the size of DNA sequences synthesized. Treatment with methylmercury hydroxide produces a partial denaturation of mRNA complexed with either itself or with other RNA molecules and results in more efficient utilization in both translational assays and DNA polymerization reactions.