Dengue Virus NS4b N-Terminus Disordered Region Interacts with NS3 Helicase C-Terminal Subdomain to Enhance Helicase Activity
Dengue virus replicates its single-stranded RNA genome in membrane-bound complexes formed on the endoplasmic reticulum, where viral non-structural proteins (NS) and RNA co-localize. The NS proteins interact with one another and with the host proteins. The interaction of the viral helicase and protea...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-08-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/8/1712 |
| _version_ | 1797407457890271232 |
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| author | Satyamurthy Kundharapu Tirumala Kumar Chowdary |
| author_facet | Satyamurthy Kundharapu Tirumala Kumar Chowdary |
| author_sort | Satyamurthy Kundharapu |
| collection | DOAJ |
| description | Dengue virus replicates its single-stranded RNA genome in membrane-bound complexes formed on the endoplasmic reticulum, where viral non-structural proteins (NS) and RNA co-localize. The NS proteins interact with one another and with the host proteins. The interaction of the viral helicase and protease, NS3, with the RNA-dependent RNA polymerase, NS5, and NS4b proteins is critical for replication. In vitro, NS3 helicase activity is enhanced by interaction with NS4b. We characterized the interaction between NS3 and NS4b and explained a possible mechanism for helicase activity modulation by NS4b. Our bacterial two-hybrid assay results showed that the N-terminal 57 residues region of NS4b is enough to interact with NS3. The molecular docking of the predicted NS4b structure onto the NS3 structure revealed that the N-terminal disordered region of NS4b wraps around the C-terminal subdomain (CTD) of the helicase. Further, NS3 helicase activity is enhanced upon interaction with NS4b. Molecular dynamics simulations on the NS4b-docked NS3 crystal structure and intrinsic tryptophan fluorescence studies suggest that the interaction results in NS3 CTD domain motions. Based on the interpretation of our results in light of the mechanism explained for NS3 helicase, NS4b–NS3 interaction modulating CTD dynamics is a plausible explanation for the helicase activity enhancement. |
| first_indexed | 2024-03-09T03:42:48Z |
| format | Article |
| id | doaj.art-bef7d22eb73e4b31b7683ddbe5c208ee |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T03:42:48Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-bef7d22eb73e4b31b7683ddbe5c208ee2023-12-03T14:39:10ZengMDPI AGViruses1999-49152022-08-01148171210.3390/v14081712Dengue Virus NS4b N-Terminus Disordered Region Interacts with NS3 Helicase C-Terminal Subdomain to Enhance Helicase ActivitySatyamurthy Kundharapu0Tirumala Kumar Chowdary1School of Biological Sciences, National Institute of Science Education and Research, An OCC of Homi Bhabha National Institute, Bhubaneswar 752050, IndiaSchool of Biological Sciences, National Institute of Science Education and Research, An OCC of Homi Bhabha National Institute, Bhubaneswar 752050, IndiaDengue virus replicates its single-stranded RNA genome in membrane-bound complexes formed on the endoplasmic reticulum, where viral non-structural proteins (NS) and RNA co-localize. The NS proteins interact with one another and with the host proteins. The interaction of the viral helicase and protease, NS3, with the RNA-dependent RNA polymerase, NS5, and NS4b proteins is critical for replication. In vitro, NS3 helicase activity is enhanced by interaction with NS4b. We characterized the interaction between NS3 and NS4b and explained a possible mechanism for helicase activity modulation by NS4b. Our bacterial two-hybrid assay results showed that the N-terminal 57 residues region of NS4b is enough to interact with NS3. The molecular docking of the predicted NS4b structure onto the NS3 structure revealed that the N-terminal disordered region of NS4b wraps around the C-terminal subdomain (CTD) of the helicase. Further, NS3 helicase activity is enhanced upon interaction with NS4b. Molecular dynamics simulations on the NS4b-docked NS3 crystal structure and intrinsic tryptophan fluorescence studies suggest that the interaction results in NS3 CTD domain motions. Based on the interpretation of our results in light of the mechanism explained for NS3 helicase, NS4b–NS3 interaction modulating CTD dynamics is a plausible explanation for the helicase activity enhancement.https://www.mdpi.com/1999-4915/14/8/1712flavivirusdengue virus (DENV)plus-stranded RNA virusviral replicationRNA helicaseDENV NS3 helicase-cum-protease |
| spellingShingle | Satyamurthy Kundharapu Tirumala Kumar Chowdary Dengue Virus NS4b N-Terminus Disordered Region Interacts with NS3 Helicase C-Terminal Subdomain to Enhance Helicase Activity Viruses flavivirus dengue virus (DENV) plus-stranded RNA virus viral replication RNA helicase DENV NS3 helicase-cum-protease |
| title | Dengue Virus NS4b N-Terminus Disordered Region Interacts with NS3 Helicase C-Terminal Subdomain to Enhance Helicase Activity |
| title_full | Dengue Virus NS4b N-Terminus Disordered Region Interacts with NS3 Helicase C-Terminal Subdomain to Enhance Helicase Activity |
| title_fullStr | Dengue Virus NS4b N-Terminus Disordered Region Interacts with NS3 Helicase C-Terminal Subdomain to Enhance Helicase Activity |
| title_full_unstemmed | Dengue Virus NS4b N-Terminus Disordered Region Interacts with NS3 Helicase C-Terminal Subdomain to Enhance Helicase Activity |
| title_short | Dengue Virus NS4b N-Terminus Disordered Region Interacts with NS3 Helicase C-Terminal Subdomain to Enhance Helicase Activity |
| title_sort | dengue virus ns4b n terminus disordered region interacts with ns3 helicase c terminal subdomain to enhance helicase activity |
| topic | flavivirus dengue virus (DENV) plus-stranded RNA virus viral replication RNA helicase DENV NS3 helicase-cum-protease |
| url | https://www.mdpi.com/1999-4915/14/8/1712 |
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