The 5′-termini of heterogeneous nuclear RNA: a comparison among molecules of different sizes and ages

The composition of the 5′ polyphosphorylated and capped termini of pulse labeled hnRNA from mouse L cells was analyzed by two-dimensional electrophoresis. Purine tri- and diphosphates: pppG, pppA, ppG and ppA; as well as four varieties of cap structure: m7GpppXm, Xm=Gm, (m6)Am, Cm and Um, were detected. With increasing labeling time the relative proportion of hnRNA molecules with tri- and diphosphorylated 5′ ends decreases and the relative proportion of capped hnRNA increases, indicating that caps are metabolically more stable than the polyphosphate termini. About half of the hnRNA molecules that are labeled within 2 hr have capped ends. This finding, together with results of earlier kinetic and structural studies, implies that a relatively high proportion of the labeled hnRNA molecules are mRNA precursors. Large hnRNA molecules exhibit a higher proportion of capped ends and a lower proportion of 5′-tri- phosphate ends as compared to small hnRNA. Given the lower stability of triphosphate termini relative to caps, this result may mean that capping of some hnRNA molecules can occur before the completion of transcription.