Characterization of the 5' termini of hn RNA in mouse L cells: implications for processing and cap formation

An analysis of the phosphorylated and capped 5' termini of the heterogenous nuclear RNA of mouse L cells has revealed four types of structure: pppXp..., ppXp..., pXp..., and m7/GpppXmp.... The 5'triphosphate termini consists exclusively of pppGp... and pppAp..., whereas a large proportion of the 5' monophosphate termini are pUp.... The 5'diphosphate termini contain all four species of nucleotide in relative proportions that are roughly similar to those found at the Xm position of cap structures. These results indicate that initiation of hnRNA transcription occurre exclusively with purine nucleotides, and consequently that the hnRNA molecules containing pyrimidines at the 5' termini very probably arise by cleavages at internal sites of larger primary transcripts. Taken together with previous results relating cap structures of hnRNA and mRNA, the data favor a model in which some mRNA sequences are located at transcriptionally initiated proportions and others in internal regions of their precursors. According to this model, both the mRNA segments derived from initial 5' end, and those derived by cleavage at internal sites could be converted to diphosphate-terminated derivatives, which then condense with GTP to form cap structures according to the mechanism previously described for vaccinia and reovirus mRNA.