Biochemical characterization of mRNA capping enzyme from Faustovirus

• DOI: 10.1261/rna.079738.123
• Publication Date: 2023-08-25T17:50:49Z
• Authors: S. Hong Chan; Christa N. Molé; Dillon Nye; Lili Mitchell; Nan Dai; Jackson Buss; Daniel W. Kneller; Joseph M. Whipple; G. Brett Robb
• Keywords: Five-prime cap; Sindbis virus

The mammalian mRNA 5′ cap structures play important roles in cellular processes such as nuclear export, efficient translation, and evading innate immune surveillance regulating 5′-mediated turnover. Hence, installation of the proper is crucial therapeutic applications synthetic mRNA. core structure, Cap-0, generated by three sequential enzymatic activities: RNA triphosphatase, guanylyltransferase, N7-guanine methyltransferase. Vaccinia virus capping enzyme (VCE) a heterodimeric that has been widely used research manufacturing. large subunit VCE D1R exhibits modular structure where each structural domains possesses one activities, whereas small D12L required to activate methyltransferase activity. Here, we report characterization single-subunit from an amoeba giant virus. Faustovirus (FCE) array catalytic common with highly generating Cap-0 without activation subunit. Phylogenetic analysis suggests FCE are descended ancestral enzyme. We found compared VCE, higher specific activity, activity toward containing secondary free end, broader temperature range, properties favorable for manufacturing workflows.