Cytoplasmic polyadenylate processing events accompany the transfer of mRNA from the free mRNP particles to the polysomes in Physarum.

Abstract

The relationship between the mRNA in the polysomes and the free cytoplasmic messenger ribonucleoprotein of Physarum polycephalum was studied by microinjection techniques. Labeled free cytoplasmic ribonucleoprotein, prepared from donor plasmodia, was microinjected into unlabeled host plasmodia, and its fat was followed in the host ribonucleoprotein particles. Approximately one-half of the poly(A)-containing RNA [poly(A)+RNA] that originated from the microinjected particles was incorporated into the host polysomes by normal translational processes within 1 hr. Very short poly(A) sequences (approximately 15 nucleotide residues) were found in these poly(A)+RNA molecules. These short poly(A) sequences were sensitive to digestion with micrococcal nuclease, suggesting that they were not associated with protein. Because the poly(A)+RNA molecules of the microinjected free cytoplasmic mRNP had originally contained poly(A) sequences 50-65 nucleotides long and were associated with protein extensive poly(A) degradation and poly(A).protein complex dissociation must have occurred during their incorporation into the polysomes or during their translation. These results demonstrate a precursor-product relationship between free cytoplasmic mRNP and polysomal mRNA and suggest that the incorporation process in Physarum is accompanied by structural modifications in the poly(A) region of mRNA. They also imply that the polysome is a site for disruption of the poly(A).protein complex and poly(A) degradation.