The addition of 5' cap structures occurs early in hnRNA synthesis and prematurely terminated molecules are capped

Abstract

After cells were labeled by brief exposure to 3H-methyl-L-methionine, the majority of labeled 5' terminal cap I (m7GpppN1mpN2p) oligonucleotide structures were in nuclear RNA (hnRNA) molecules approximately 750 nucleotides or less in length. After longer label times, the proportion of cap I structures in nuclear molecules longer than mRNA rose to approximately 60% of the total, but approximately 40% of the cap I structures were still in molecules shorter than approximately 750 nucleotides. The cap I structures in both long and short hnRNA chains contained all four 2' methylated nucleotides in the N1 position in about the same proportion as in mRNA. None of the large hnRNA molecules could be demonstrated to contain 5' pppX p termini; the only such terminus in high molecular weight RNA was pppAp which was decreased markedly by low doses of actinomycin and is presumably the terminus of pre-rRNA. These results raise the possibilities that hnRNA chains can initiate with any of the four nucleotides, that capping occurs very close to or at the start of hnRNA chain synthesis and that approximately 40% of the hnRNA chains may be prematurely terminated.