The immunosuppressive agent mizoribine monophosphate is an inhibitor of the human RNA capping enzyme.
Mizoribine monophosphate (MZP) is a specific inhibitor of the cellular inosine-5'-monophosphate dehydrogenase (IMPDH), the enzyme catalyzing the rate-limiting step of de novo guanine nucleotide biosynthesis. MZP is a highly potent antagonistic inhibitor of IMPDH that blocks the proliferation of T and B lymphocytes that use the de novo pathway of guanine nucleotide synthesis almost exclusively. In the present study, we investigated the ability of MZP to directly inhibit the human RNA capping enzyme (HCE), a protein harboring both RNA 5'-triphosphatase and RNA guanylyltransferase activities. HCE is involved in the synthesis of the cap structure found at the 5' end of eukaryotic mRNAs, which is critical for the splicing of the cap-proximal intron, the transport of mRNAs from the nucleus to the cytoplasm, and for both the stability and translation of mRNAs. Our biochemical studies provide the first insight that MZP can inhibit the formation of the RNA cap structure catalyzed by HCE. In the presence of MZP, the RNA 5'-triphosphatase activity appears to be relatively unaffected while the RNA guanylyltransferase activity is inhibited, indicating that the RNA guanylyltransferase activity is the main target of MZP inhibition. Kinetic studies reveal that MZP is a non-competitive inhibitor that likely targets an allosteric site on HCE. Mizoribine also impairs mRNA capping in living cells, which could account for the global mechanism of action of this therapeutic agent. Together, our study clearly demonstrates that mizoribine monophosphate inhibits the human RNA guanylyltransferase in vitro and impair mRNA capping in cellulo.
Département de Biochimie, Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, Sherbrooke, Québec, Canada.
Frédéric
F
Picard-Jean
Isabelle
I
Bougie
Satoshi
S
Shuto
Martin
M
Bisaillon
D017766:Acid Anhydride Hydrolases / Q000737:chemistry; D001402:B-Lymphocytes / Q000201:enzymology; D002384:Catalysis; D006150:Guanine Nucleotides / Q000096:biosynthesis; D057809:HEK293 Cells; D006801:Humans; D007166:Immunosuppressive Agents / Q000008:administration & dosage*; D007700:Kinetics; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics*; D012315:RNA Caps / Q000737:chemistry / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D012263:Ribonucleosides / Q000008:administration & dosage* / Q000737:chemistry; D013601:T-Lymphocytes / Q000201:enzymology; D000098556:mRNA Guanylyltransferases
D006150:Guanine Nucleotides; D007166:Immunosuppressive Agents; D012315:RNA Caps; D012333:RNA, Messenger; D012263:Ribonucleosides; C010052:mizoribine; D009713:Nucleotidyltransferases; D000098556:mRNA Guanylyltransferases; D017766:Acid Anhydride Hydrolases; C023090:RNA triphosphatase