Structural and mutagenesis studies of the NS3 helicase from dengue virus for drug discovery

With reference to the structure analysis of RNA unwinding mechanism coupled with ATP hydrolysis by Dengue virus (DEN) non-structural 3 (NS3) protein (Luo et al., 2008a), site-directed mutagenesis on DEN NS3 helicase domain was performed. Five residues conserved among Flaviviridae were selected for m...

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Main Author: Yang, Xiaoqing
Other Authors: Julien Lescar
Format: Final Year Project
Language:English
Published: 2009
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Online Access:http://hdl.handle.net/10356/18935
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-189352023-02-28T18:00:21Z Structural and mutagenesis studies of the NS3 helicase from dengue virus for drug discovery Yang, Xiaoqing Julien Lescar School of Biological Sciences DRNTU::Science::Biological sciences::Microbiology::Virology With reference to the structure analysis of RNA unwinding mechanism coupled with ATP hydrolysis by Dengue virus (DEN) non-structural 3 (NS3) protein (Luo et al., 2008a), site-directed mutagenesis on DEN NS3 helicase domain was performed. Five residues conserved among Flaviviridae were selected for mutation and six soluble mutant proteins were generated. RNA stimulated ATPase activity and RNA helicase activity were determined. Residue D290 resides in subdomain I and is able to form a direct contact to RNA. Mutation D290A presented significantly reduced ATPase activity and enhanced helicase activity. Residue L429 resides in a F hairpin of subdomain II. It pairs with A444 to form a hydrophobic patch, which was proposed to disrupt base stacking at RNA unwinding fork. Mutation L429V enhanced helicase activity. A455 resides in VI motif at subdomain II and was suggested to form a part of phosphate exit route. Mutations A455I and A455T diminished ATP hydrolysis activity. Protein activities were further discussed in a structure-based manner. The study might be useful for future investigation on DEN NS3 and antiviral drug design. 2009-08-21T07:05:46Z 2009-08-21T07:05:46Z 2009 2009 Final Year Project (FYP) http://hdl.handle.net/10356/18935 en Nanyang Technological University 40 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Biological sciences::Microbiology::Virology
spellingShingle DRNTU::Science::Biological sciences::Microbiology::Virology
Yang, Xiaoqing
Structural and mutagenesis studies of the NS3 helicase from dengue virus for drug discovery
description With reference to the structure analysis of RNA unwinding mechanism coupled with ATP hydrolysis by Dengue virus (DEN) non-structural 3 (NS3) protein (Luo et al., 2008a), site-directed mutagenesis on DEN NS3 helicase domain was performed. Five residues conserved among Flaviviridae were selected for mutation and six soluble mutant proteins were generated. RNA stimulated ATPase activity and RNA helicase activity were determined. Residue D290 resides in subdomain I and is able to form a direct contact to RNA. Mutation D290A presented significantly reduced ATPase activity and enhanced helicase activity. Residue L429 resides in a F hairpin of subdomain II. It pairs with A444 to form a hydrophobic patch, which was proposed to disrupt base stacking at RNA unwinding fork. Mutation L429V enhanced helicase activity. A455 resides in VI motif at subdomain II and was suggested to form a part of phosphate exit route. Mutations A455I and A455T diminished ATP hydrolysis activity. Protein activities were further discussed in a structure-based manner. The study might be useful for future investigation on DEN NS3 and antiviral drug design.
author2 Julien Lescar
author_facet Julien Lescar
Yang, Xiaoqing
format Final Year Project
author Yang, Xiaoqing
author_sort Yang, Xiaoqing
title Structural and mutagenesis studies of the NS3 helicase from dengue virus for drug discovery
title_short Structural and mutagenesis studies of the NS3 helicase from dengue virus for drug discovery
title_full Structural and mutagenesis studies of the NS3 helicase from dengue virus for drug discovery
title_fullStr Structural and mutagenesis studies of the NS3 helicase from dengue virus for drug discovery
title_full_unstemmed Structural and mutagenesis studies of the NS3 helicase from dengue virus for drug discovery
title_sort structural and mutagenesis studies of the ns3 helicase from dengue virus for drug discovery
publishDate 2009
url http://hdl.handle.net/10356/18935
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