Purification of mRNA guanylyltransferase from vaccinia virions.

Abstract

GTP:RNA guanylyltransferase, the enzyme which catalyzes the guanylylation of the 5' termini of viral mRNAs, has been isolated and purified approximately 10,000-fold from cores of vaccinia virus. S-adenosyl-methionine:mRNA (guanine-7)-methyltransferase copurified with guanylyltransferase activity through chromatography on DNA agarose, phosphocellulose, and centrifugation in glycerol gradients, suggesting that the two activities are closely associated. The molecular weight of native guanylyltransferase- and 7-methyltransferase-associated activities was approximately 120,000 as determined by glycerol gradient centrifugation. Guanylytransferase purified by electrophoresis on polyacrylamide gels at pH 4.5 lacked 7-methyltransferase activity. Analysis by electrophoresis on sodium dodecyl sulfate-polyacrylamide gels of electrophoretically purified native guanylyltransferase showed the presence of one major band of polypeptide which had a molecular weight of approximately 59,000.

• Authors: G Monroy, E Spencer, J Hurwitz
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 253 Issue 12 Pg. 4481-9 (Jun 25 1978) ISSN: 0021-9258 [Print] United States
• PMID: 659428 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Guanine Nucleotides
• RNA, Messenger
• Methyltransferases
• Nucleotidyltransferases
• mRNA guanylyltransferase

Topics

• Guanine Nucleotides
• Methyltransferases (isolation & purification, metabolism)
• Nucleotidyltransferases (isolation & purification, metabolism)
• RNA, Messenger
• Vaccinia virus (enzymology)
• Virion (enzymology)