title: Quantitative assessment of mRNA cap analogues as inhibitors of in vitro translation.
abstract: Fifty-eight analogues of the 5'-terminal 7-methylguanosine-containing cap of eukaryotic messenger RNA were synthesized and tested for their ability to inhibit in vitro protein synthesis. A new algorithm was developed for extracting KI, the dissociation constant for the cap analogue.eIF4E complex, from protein synthesis data. The results indicated that addition of a methyl group to the N2 of guanine produced more inhibitory compounds, but addition of a second methyl group to N2 decreased the level of inhibition dramatically. Aryl substitution at N7 improved the efficacy of guanine nucleoside monophosphate analogues. Substitution of the aromatic ring at the para position with methyl or NO2 groups abolished this effect, but substitution with Cl or F enhanced it. By contrast, aryl substitution at N7 in nucleoside di- or triphosphate analogues produced only minor effects, both positive and negative. By far the strongest determinants of inhibitory activity for cap analogues were phosphate residues. The beneficial effect of more phosphate residues was related more to anionic charge than to the number of phosphate groups per se. The second nucleotide residue in analogues of the form m7GpppN affected inhibitory activity in the order G > C > U > A, but there was no effect of 2'-O-modification. Opening the first ribose ring of m7GpppG analogues dramatically decreased activity, but alterations at the 2'-position of this ribose had no effect. Non-nucleotide-based cap analogues containing benzimidazole derivatives were inhibitory, though less so than those containing 7-methylguanine.
authors.affiliation: Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, Shreveport 71130-3932, USA.
authors.forename: A
authors.initials: A
authors.lastname: Cai
authors.forename: M
authors.initials: M
authors.lastname: Jankowska-Anyszka
authors.forename: A
authors.initials: A
authors.lastname: Centers
authors.forename: L
authors.initials: L
authors.lastname: Chlebicka
authors.forename: J
authors.initials: J
authors.lastname: Stepinski
authors.forename: R
authors.initials: R
authors.lastname: Stolarski
authors.forename: E
authors.initials: E
authors.lastname: Darzynkiewicz
authors.forename: R E
authors.initials: RE
authors.lastname: Rhoads
mesh_terms: D000818:Animals; D001562:Benzimidazoles / Q000737:chemistry; D002723:Chlorobenzoates / Q000737:chemistry; D039561:Eukaryotic Initiation Factor-4E; D007700:Kinetics; D010448:Peptide Initiation Factors / Q000037:antagonists & inhibitors / Q000737:chemistry; D010710:Phosphates / Q000737:chemistry; D014176:Protein Biosynthesis / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D012314:RNA Cap Analogs / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D012333:RNA, Messenger / Q000138:chemical synthesis / Q000494:pharmacology; D011817:Rabbits; D012266:Ribose / Q000737:chemistry
chemical_list: D001562:Benzimidazoles; D002723:Chlorobenzoates; D039561:Eukaryotic Initiation Factor-4E; D010448:Peptide Initiation Factors; D010710:Phosphates; D011500:Protein Synthesis Inhibitors; D012314:RNA Cap Analogs; D012333:RNA, Messenger; C011925:7-methylguanosine-5'-monophosphate; D012266:Ribose; C031000:benzimidazole; C040768:4-chlorobenzoic acid