Polysome Analysis

During the cyclic process of translation, a small (40S) and large (60S) ribosomal subunit associate with mRNA to form an 80S complex (monosome). This ribosome moves along the mRNA during translational elongation, and then dissociates into the 40S and 60S subunits on termination. During elongation by one ribosome, further ribosomes can initiate translation on the same mRNA to form polysomes. Each polysomal complex can contain from two to over twenty ribosomes, and the mass of each complex is determined primarily by the number of ribosomes it contains. Hence, the population of polysomes within the cell can be size-fractionated by sucrose density gradient centrifugation on the basis of the loading of ribosomes on the mRNA. Also, mRNA that is being actively translated can be fractionated from untranslated mRNA by separating polysomal and monosomal material by centrifugation through sucrose shelves.