Biomedical subjects
Publications and source records attributed to D E Croall.
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Polysomal RNAs were isolated from control neuroblastoma cells and those treated with 1,N6-dibutyrl-adenosine 3',5'-phosphate (Bt2cAMP) and translated in wheat germ lysates. Comparison of proteins synthesized in vitro on two-dimensional gel electrophoretograms showed that there was a specific induction in the synthesis of a protein, Mr 48000, by the polysomal RNAs from Bt2cAMP-treated cells. This protein was identified as the R1 cAMP-binding protein by its coelectrophoresis with unlabelled binding protein and by its specific retention on 8-(6-aminohexylamino)-adenosine 3',5'-phosphate linked to Sepharose. Quantification of the proteins synthesized in vitro with subsaturating inputs of polysomal RNAs showed that there was a 1.4--1.7-fold increase in the synthesis of the R1 cAMP-binding protein by polysomal RNAs isolated from Bt2cAMP-treated cells. There was a similar increase when purified polyadenylated mRNA populations were compared. showing there was no change in the ratio of adenylated to nonadenylated mRNAs in the induced mRNA population. There was no corresponding increase in the synthesis of the R2 cAMP-binding protein although the relative synthesis of several other proteins was also increased and the synthesis of actin and the alpha and beta-tubulin subunits was decreased. The increased levels of the R1 cAMP-binding protein found in Bt2cAMP-treated neuroblastoma cells are therefore partly caused by a specific accumulation of its mRNA on polysomes. The mRNA content of the cytoplasmic messenger ribonucleoprotein (mRNP) population of control cells was insufficient to account for this increase by a translocation of R1 mRNA from the mRNP to the polysome fraction in Bt2cAMP-treated cells. The increase in polysomal R1 mRNA is therefore caused by its increased transcription of post-transcriptional processing or its decreased rate of degradation in Bt2cAMP-treated cells. Although the R1 and R2 binding proteins have identical molecular weights and similar pI values, the specific induction of the mRNA for R1 cAMP-binding protein and the differential distribution of the R1 and R2 mRNAs between the polysomal and messenger ribonucleoprotein compartments show that these two cAMP-binding proteins are encoded by different mRNA populations.
Highly purified poly(A)-containing free and polysomal mRNP particles have been isolated by chromatography of subcellular fractions of chick embryonic muscles on oligo-dT-cellulose and elution with low salt buffer at 45 degrees. The free and polysomal mRNP represent two distinct classes of macromolecules, the free particles having a more complex nucleoprotein organization than the polysomal particles. Comparison of the protein moieties of three classes of poly(A)-containing cytoplasmic mRNP -- those released from nuclei after in vitro transcription and processing (transported mRNP), the free, and polysomal mRNP -- strongly suggests that the majority of the mRNA-associated proteins are exchanged in the cytoplasm during the various functional states of mRNA. A model of translational control involving the participation of mRNA-associated proteins in chick embryonic muscles and by analogy in other differentiated eukaryotic cells is proposed.