Dual modes of RNA-silencing suppression by Flock House virus protein B2

As a counter-defense against antiviral RNA silencing during infection, the insect Flock House virus (FHV) expresses the silencing suppressor protein B2. Biochemical experiments show that B2 binds to double-stranded RNA (dsRNA) without regard to length and inhibits cleavage of dsRNA by Dicer in vitro. A cocrystal structure reveals that a B2 dimer forms a four-helix bundle that binds to one face of an A-form RNA duplex independently of sequence. These results suggest that B2 blocks both cleavage of the FHV genome by Dicer and incorporation of FHV small interfering RNAs into the RNA-induced silencing complex.

[1]  Morris F. Maduro,et al.  Animal virus replication and RNAi-mediated antiviral silencing in Caenorhabditis elegans , 2005, Nature.

[2]  Zsuzsanna Lichner,et al.  Double-stranded RNA-binding proteins could suppress RNA interference-mediated antiviral defences. , 2003, The Journal of general virology.

[3]  G. Winter,et al.  Identification of functional similarities between proteins using directed evolution , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[4]  M Kokkinidis,et al.  Structure of the ColE1 rop protein at 1.7 A resolution. , 1987, Journal of molecular biology.

[5]  L. Malinina,et al.  Recognition of small interfering RNA by a viral suppressor of RNA silencing , 2003, Nature.

[6]  J. Williamson,et al.  Central domain assembly: thermodynamics and kinetics of S6 and S18 binding to an S15-RNA complex. , 2001, Journal of molecular biology.

[7]  Thomas C. Terwilliger,et al.  Automated MAD and MIR structure solution , 1999, Acta crystallographica. Section D, Biological crystallography.

[8]  S. Doublié Preparation of selenomethionyl proteins for phase determination. , 1997, Methods in enzymology.

[9]  A. Lupas Prediction and analysis of coiled-coil structures. , 1996, Methods in enzymology.

[10]  Olivier Voinnet,et al.  Induction and suppression of RNA silencing: insights from viral infections , 2005, Nature Reviews Genetics.

[11]  Robert M. Goodman,et al.  Systemic spread of an RNA insect virus in plants expressing plant viral movement protein genes , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[12]  Kyle L. Johnson,et al.  Nodamura Virus Nonstructural Protein B2 Can Enhance Viral RNA Accumulation in both Mammalian and Insect Cells , 2004, Journal of Virology.

[13]  Tom Alber,et al.  Automated protein crystal structure determination using ELVES. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[15]  A. Lupas,et al.  Predicting coiled coils from protein sequences , 1991, Science.

[16]  N L Harris,et al.  Four helix bundle diversity in globular proteins. , 1994, Journal of molecular biology.

[17]  B. Jacobs,et al.  Both Carboxy- and Amino-Terminal Domains of the Vaccinia Virus Interferon Resistance Gene, E3L, Are Required for Pathogenesis in a Mouse Model , 2001, Journal of Virology.

[18]  F. Crick,et al.  The packing of α‐helices: simple coiled‐coils , 1953 .

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

[20]  J. Thornton,et al.  PROCHECK: a program to check the stereochemical quality of protein structures , 1993 .

[21]  I. Tinoco,et al.  Mapping of a protein-RNA kissing hairpin interface: Rom and Tar-Tar*. , 1998, Nucleic acids research.

[22]  R. Rueckert,et al.  Black Beetle Virus: Messenger for Protein B Is a Subgenomic Viral RNA , 1982, Journal of virology.

[23]  L. A. Ball,et al.  Comparisons among the larger genome segments of six nodaviruses and their encoded RNA replicases. , 2001, The Journal of general virology.

[24]  D E McRee,et al.  XtalView/Xfit--A versatile program for manipulating atomic coordinates and electron density. , 1999, Journal of structural biology.

[25]  S C Schultz,et al.  Molecular basis of double‐stranded RNA‐protein interactions: structure of a dsRNA‐binding domain complexed with dsRNA , 1998, The EMBO journal.

[26]  G. Szittya,et al.  Molecular mechanism of RNA silencing suppression mediated by p19 protein of tombusviruses , 2004, The EMBO journal.

[27]  É. Várallyay,et al.  Plant Virus-Derived Small Interfering RNAs Originate Predominantly from Highly Structured Single-Stranded Viral RNAs , 2005, Journal of Virology.

[28]  J. Tomizawa,et al.  Control of cole 1 plasmid replication: Enhancement of binding of RNA I to the primer transcript by the rom protein , 1984, Cell.

[29]  S. Ding,et al.  Induction and Suppression of RNA Silencing by an Animal Virus , 2002, Science.

[30]  R Lavery,et al.  The definition of generalized helicoidal parameters and of axis curvature for irregular nucleic acids. , 1988, Journal of biomolecular structure & dynamics.

[31]  Chris Sander,et al.  Dali/FSSP classification of three-dimensional protein folds , 1997, Nucleic Acids Res..

[32]  Thomas C. Terwilliger,et al.  Electronic Reprint Biological Crystallography Maximum-likelihood Density Modification , 2022 .

[33]  D. Ganem,et al.  A Virus-Encoded Inhibitor That Blocks RNA Interference in Mammalian Cells , 2005, Journal of Virology.

[34]  L. Regan,et al.  Dissecting RNA-protein interactions: RNA-RNA recognition by Rop , 1995, Cell.

[35]  Shou-Wei Ding,et al.  Interferon antagonist proteins of influenza and vaccinia viruses are suppressors of RNA silencing. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[36]  G. Szittya,et al.  Size Selective Recognition of siRNA by an RNA Silencing Suppressor , 2003, Cell.

[37]  James M Aramini,et al.  Biophysical characterization of the complex between double-stranded RNA and the N-terminal domain of the NS1 protein from influenza A virus: evidence for a novel RNA-binding mode. , 2004, Biochemistry.