RNA capping by the vaccinia virus guanylyltransferase. Structure of enzyme-guanylate intermediate.
GTP:RNA guanylyltransferase isolated from vaccinia virus catalyzes the transfer of GMP from GTP to the 5' terminus of RNA via an enzyme-guanylate intermediate. Incubation of the purified vaccinia RNA guanylyltransferase with [alpha- 32P]GTP and MgCl2 yields [32P]GMP covalently linked to the Mr = 95,000 subunit. The bond involves the phosphate moiety of GMP and the Ne-amino group of lysine. This was verified by treatment of the isolated 95-kDa subunit-[32P]GMP complex with sodium periodate, followed by methylamine-catalyzed beta-elimination. The product was then hydrolyzed by alkali producing 32P-labeled lysine (Ne-P)phosphate.
D010450:Endopeptidases; D006160:Guanosine Triphosphate / Q000378:metabolism; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D008970:Molecular Weight; D009713:Nucleotidyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis; D011485:Protein Binding; D012315:RNA Caps / Q000378:metabolism*; D014616:Vaccinia virus / Q000201:enzymology*
D010446:Peptide Fragments; D012315:RNA Caps; D006160:Guanosine Triphosphate; D009713:Nucleotidyltransferases; C020842:guanylyltransferase; D010450:Endopeptidases