Small Molecule Inhibitors That Selectively Block Dengue Virus Methyltransferase*

Crystal structure analysis of Flavivirus methyltransferases uncovered a flavivirus-conserved cavity located next to the binding site for its cofactor, S-adenosyl-methionine (SAM). Chemical derivatization of S-adenosyl-homocysteine (SAH), the product inhibitor of the methylation reaction, with substituents that extend into the identified cavity, generated inhibitors that showed improved and selective activity against dengue virus methyltransferase (MTase), but not related human enzymes. Crystal structure of dengue virus MTase with a bound SAH derivative revealed that its N6-substituent bound in this cavity and induced conformation changes in residues lining the pocket. These findings demonstrate that one of the major hurdles for the development of methyltransferase-based therapeutics, namely selectivity for disease-related methyltransferases, can be overcome.

[1]  C. Lima,et al.  Structure and mechanism of mRNA cap (guanine-N7) methyltransferase. , 2004, Molecular cell.

[2]  P. Shi,et al.  Characterization of Dengue Virus Resistance to Brequinar in Cell Culture , 2010, Antimicrobial Agents and Chemotherapy.

[3]  S. Vasudevan,et al.  Mutagenesis of the Dengue Virus Type 2 NS5 Methyltransferase Domain* , 2008, Journal of Biological Chemistry.

[4]  A G Leslie,et al.  Biological Crystallography Integration of Macromolecular Diffraction Data , 2022 .

[5]  Kevin Cowtan,et al.  research papers Acta Crystallographica Section D Biological , 2005 .

[6]  S. Jacobsen,et al.  Substrate Specificity and Kinetic Mechanism of Mammalian G9a Histone H3 Methyltransferase* , 2004, Journal of Biological Chemistry.

[7]  R. Borchardt,et al.  Relationship between intracellular concentration of S-adenosylhomocysteine and inhibition of vaccinia virus replication and inhibition of murine L-929 cell growth , 1989, Antimicrobial Agents and Chemotherapy.

[8]  Yi Guo,et al.  West Nile Virus 5′-Cap Structure Is Formed by Sequential Guanine N-7 and Ribose 2′-O Methylations by Nonstructural Protein 5 , 2006, Journal of Virology.

[9]  Y. Cheng,et al.  Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction. , 1973, Biochemical pharmacology.

[10]  Yi Guo,et al.  Structure and Function of Flavivirus NS5 Methyltransferase , 2007, Journal of Virology.

[11]  P. Shi,et al.  Structural and Functional Analyses of a Conserved Hydrophobic Pocket of Flavivirus Methyltransferase* , 2010, The Journal of Biological Chemistry.

[12]  A. Thompson,et al.  Analysis of flavivirus NS5 methyltransferase cap binding. , 2009, Journal of molecular biology.

[13]  David I. Stuart,et al.  Structure and functionality in flavivirus NS-proteins: Perspectives for drug design , 2010, Antiviral research.

[14]  Robert A. Copeland,et al.  Protein methyltransferases as a target class for drug discovery , 2009, Nature Reviews Drug Discovery.

[15]  Nadine H. Elowe,et al.  SAR around (l)-S-adenosyl-l-homocysteine, an inhibitor of human DNA methyltransferase (DNMT) enzymes. , 2009, Bioorganic & medicinal chemistry letters.

[16]  F. Quiocho,et al.  The 1.85 Å Structure of Vaccinia Protein VP39: A Bifunctional Enzyme That Participates in the Modification of Both mRNA Ends , 1996, Cell.

[17]  Hongping Dong,et al.  Biochemical and genetic characterization of dengue virus methyltransferase. , 2010, Virology.

[18]  S J Wodak,et al.  SFCHECK: a unified set of procedures for evaluating the quality of macromolecular structure-factor data and their agreement with the atomic model. , 1999, Acta crystallographica. Section D, Biological crystallography.

[19]  Jean-Louis Romette,et al.  An RNA cap (nucleoside‐2′‐O‐)‐methyltransferase in the flavivirus RNA polymerase NS5: crystal structure and functional characterization , 2002, The EMBO journal.

[20]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[21]  D. Wen,et al.  A scintillation proximity assay for dengue virus NS5 2'-O-methyltransferase-kinetic and inhibition analyses. , 2008, Antiviral research.

[22]  R. Borchardt.  S-Adenosyl-L-methionine-dependent macromolecule methyltransferases: potential targets for the design of chemotherapeutic agents. , 1980, Journal of medicinal chemistry.

[23]  P. Shi,et al.  Higher catalytic efficiency of N-7-methylation is responsible for processive N-7 and 2'-O methyltransferase activity in dengue virus. , 2010, Virology.