7t7h
From Proteopedia
Crystal structure of Vaccinia Virus decapping enzyme D9 in complex with inhibitor CP100356
Structural highlights
Function[L7QJE0_9POXV] Decapping enzyme required for the removal of the 5'-end m7GpppN cap tethered to viral and host mRNAs to allow their decay in cells. May therefore accelerate viral and cellular mRNA turnover to eliminate competing host mRNAs and allow stage-specific synthesis of viral proteins. Acceleration of the turnover of cellular transcripts may even promote the shutoff of host protein synthesis. Does not cleave unmethylated RNAs or RNAs shorter than 24 nucleotides.[ARBA:ARBA00025519] Publication Abstract from PubMedVaccinia virus (VACV) represents a family of poxviruses, which possess their own decapping machinery as a part of their strategy to eliminate host mRNAs and evade the innate immune response. D9 is one of the two encoded VACV decapping enzymes that is responsible for cap removal from the 5' end of both host mRNA transcripts and viral double-stranded RNAs. Little is known about the structural requirements for D9 inhibition by small molecules. Here, we identified a minimal D9 substrate and used it to develop a real-time fluorescence assay for inhibitor discovery and characterization. We screened a panel of nucleotide-derived substrate analogues and pharmacologically active candidates to identify several compounds with nano- and low micromolar IC50 values. m(7)GpppCH2p was the most potent nucleotide inhibitor (IC50 approximately 0.08 muM), and seliciclib and CP-100356 were the most potent drug-like compounds (IC50 0.57 and 2.7 muM, respectively). The hits identified through screening inhibited D9-catalyzed decapping of 26 nt RNA substrates but were not active toward VACV D10 or human decapping enzyme, Dcp1/2. The inhibition mode for one of the compounds (CP-100356) was elucidated based on the X-ray cocrystal structure, opening the possibility for structure-based design of novel D9 inhibitors and binding probes. Fluorescence-Based Activity Screening Assay Reveals Small Molecule Inhibitors of Vaccinia Virus mRNA Decapping Enzyme D9.,Bednarczyk M, Peters JK, Kasprzyk R, Starek J, Warminski M, Spiewla T, Mugridge JS, Gross JD, Jemielity J, Kowalska J ACS Chem Biol. 2022 May 16. doi: 10.1021/acschembio.2c00049. PMID:35576528[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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