5'-Terminal capping of RNA by guanylyltransferase from HeLa cell nuclei

Abstract

A soluble extract prepared from HeLa cell nuclei has been shown to catalyze the 5'-terminal modification of RNA and synthetic polyribonucleotides to form m7G(5')pppA-and m7G(5')-pppG- structures referred to as caps. The reaction involves the transfer of a GMP moiety from GTP to the 5' end of an RNA molecule containing at least two terminal phosphates. Significantly, neither the beta nor the gamma phosphates of GTP are transferred and polynucleotides with no 5'-terminal phosphate or only one are not acceptors. In the absence of methyl donor, G(5')pppA- and G(5')pppG- structures were synthesized, indicating that methylation is not required for guanylylation. Cap formation was considered to occur by the following mechanism: (see article), in which AdoMet is S-adenosylmethionine, AdoHcy is S-adenosylhomocysteine, and (p)ppN- represents either the original 5' end of an RNA molecule or an internal site to which one or more phosphates were added after processing.