Structure and Function of Flavivirus NS5 Methyltransferase

Zhou, Y; Ray, D; Zhao, Y; Dong, H; Ren, S; Li, Z; Guo, Y; Bernard, K; Shi, P; Li, H

doi:10.1128/JVI.02704-06

Title: Structure and Function of Flavivirus NS5 Methyltransferase

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Abstract

The plus-strand RNA genome of flavivirus contains a 5' terminal cap 1 structure (m{sup 7}GpppAmG). The flaviviruses encode one methyltransferase, located at the N-terminal portion of the NS5 protein, to catalyze both guanine N-7 and ribose 2'-OH methylations during viral cap formation. Representative flavivirus methyltransferases from dengue, yellow fever, and West Nile virus (WNV) sequentially generate GpppA {yields} m{sup 7}GpppA {yields} m{sup 7}GpppAm. The 2'-O methylation can be uncoupled from the N-7 methylation, since m{sup 7}GpppA-RNA can be readily methylated to m{sup 7}GpppAm-RNA. Despite exhibiting two distinct methylation activities, the crystal structure of WNV methyltransferase at 2.8 {angstrom} resolution showed a single binding site for S-adenosyl-L-methionine (SAM), the methyl donor. Therefore, substrate GpppA-RNA should be repositioned to accept the N-7 and 2'-O methyl groups from SAM during the sequential reactions. Electrostatic analysis of the WNV methyltransferase structure showed that, adjacent to the SAM-binding pocket, is a highly positively charged surface that could serve as an RNA binding site during cap methylations. Biochemical and mutagenesis analyses show that the N-7 and 2'-O cap methylations require distinct buffer conditions and different side chains within the K{sub 61}-D{sub 146}-K{sub 182}-E{sub 218} motif, suggesting that the two reactions use different mechanisms. In the contextmore »« less

Authors:: Zhou, Y; Ray, D; Zhao, Y; Dong, H; Ren, S; Li, Z; Guo, Y; Bernard, K; Shi, P; Li, H

Publication Date:: Mon Jan 01 00:00:00 EST 2007

Research Org.:: Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source

Sponsoring Org.:: Doe - Office Of Science

OSTI Identifier:: 930027

Report Number(s):: BNL-80641-2008-JA
Journal ID: ISSN 0022-538X; JOVIAM; TRN: US200822%%1263

DOE Contract Number:: DE-AC02-98CH10886

Resource Type:: Journal Article

Journal Name:: Journal of Virology

Additional Journal Information:: Journal Volume: 81; Journal ID: ISSN 0022-538X

Country of Publication:: United States

Language:: English

Subject:: 36 MATERIALS SCIENCE; BUFFERS; CRYSTAL STRUCTURE; DEFECTS; ELECTROSTATICS; FEVER; FUNCTIONS; GUANINE; LETHAL MUTATIONS; LIFE CYCLE; METHYLATION; MICE; MUTAGENESIS; RESOLUTION; RIBOSE; RNA; SUBSTRATES; SURFACES; THERAPY; VIRUSES; national synchrotron light source

Citation Formats


                    Zhou, Y, Ray, D, Zhao, Y, Dong, H, Ren, S, Li, Z, Guo, Y, Bernard, K, Shi, P, and Li, H. Structure and Function of Flavivirus NS5 Methyltransferase.  United States: N. p., 2007. 
        Web.  doi:10.1128/JVI.02704-06.

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                    Zhou, Y, Ray, D, Zhao, Y, Dong, H, Ren, S, Li, Z, Guo, Y, Bernard, K, Shi, P, & Li, H. Structure and Function of Flavivirus NS5 Methyltransferase.  United States.  https://doi.org/10.1128/JVI.02704-06

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                    Zhou, Y, Ray, D, Zhao, Y, Dong, H, Ren, S, Li, Z, Guo, Y, Bernard, K, Shi, P, and Li, H. 2007.  
        "Structure and Function of Flavivirus NS5 Methyltransferase".  United States.  https://doi.org/10.1128/JVI.02704-06.

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@article{osti_930027,

  title        = {Structure and Function of Flavivirus NS5 Methyltransferase},

  author       = {Zhou, Y and Ray, D and Zhao, Y and Dong, H and Ren, S and Li, Z and Guo, Y and Bernard, K and Shi, P and Li, H},

  abstractNote = {The plus-strand RNA genome of flavivirus contains a 5' terminal cap 1 structure (m{sup 7}GpppAmG). The flaviviruses encode one methyltransferase, located at the N-terminal portion of the NS5 protein, to catalyze both guanine N-7 and ribose 2'-OH methylations during viral cap formation. Representative flavivirus methyltransferases from dengue, yellow fever, and West Nile virus (WNV) sequentially generate GpppA {yields} m{sup 7}GpppA {yields} m{sup 7}GpppAm. The 2'-O methylation can be uncoupled from the N-7 methylation, since m{sup 7}GpppA-RNA can be readily methylated to m{sup 7}GpppAm-RNA. Despite exhibiting two distinct methylation activities, the crystal structure of WNV methyltransferase at 2.8 {angstrom} resolution showed a single binding site for S-adenosyl-L-methionine (SAM), the methyl donor. Therefore, substrate GpppA-RNA should be repositioned to accept the N-7 and 2'-O methyl groups from SAM during the sequential reactions. Electrostatic analysis of the WNV methyltransferase structure showed that, adjacent to the SAM-binding pocket, is a highly positively charged surface that could serve as an RNA binding site during cap methylations. Biochemical and mutagenesis analyses show that the N-7 and 2'-O cap methylations require distinct buffer conditions and different side chains within the K{sub 61}-D{sub 146}-K{sub 182}-E{sub 218} motif, suggesting that the two reactions use different mechanisms. In the context of complete virus, defects in both methylations are lethal to WNV; however, viruses defective solely in 2'-O methylation are attenuated and can protect mice from later wild-type WNV challenge. The results demonstrate that the N-7 methylation activity is essential for the WNV life cycle and, thus, methyltransferase represents a novel target for flavivirus therapy.},

  doi          = {10.1128/JVI.02704-06},

  url          = {https://www.osti.gov/biblio/930027},
  journal      = {Journal of Virology},

  issn         = {0022-538X},

  number       = ,

  volume       = 81,

  place        = {United States},

  year         = {Mon Jan 01 00:00:00 EST 2007},

  month        = {Mon Jan 01 00:00:00 EST 2007}

}

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