Saddle curvature association of nsP1 facilitates the replication complex assembly of Chikungunya virus in cells

Abstract Positive-sense RNA viruses, including SARS-CoV-1 and -2, DENV, and CHIKV, replicate in curved membrane compartments within host cells. Non-structural proteins (nsPs) critically regulate these nanoscale membrane structures, yet their curvature-dependent assembly remains elusive due to the ch...

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Main Authors: Xinwen Miao, Michelle Cheok Yien Law, Jatin Kumar, Choon-Peng Chng, Yongpeng Zeng, Yaw Bia Tan, Jiawei Wu, Xiangfu Guo, Lizhen Huang, Yinyin Zhuang, Weibo Gao, Changjin Huang, Dahai Luo, Wenting Zhao
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-59402-0
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author Xinwen Miao
Michelle Cheok Yien Law
Jatin Kumar
Choon-Peng Chng
Yongpeng Zeng
Yaw Bia Tan
Jiawei Wu
Xiangfu Guo
Lizhen Huang
Yinyin Zhuang
Weibo Gao
Changjin Huang
Dahai Luo
Wenting Zhao
author_facet Xinwen Miao
Michelle Cheok Yien Law
Jatin Kumar
Choon-Peng Chng
Yongpeng Zeng
Yaw Bia Tan
Jiawei Wu
Xiangfu Guo
Lizhen Huang
Yinyin Zhuang
Weibo Gao
Changjin Huang
Dahai Luo
Wenting Zhao
author_sort Xinwen Miao
collection DOAJ
description Abstract Positive-sense RNA viruses, including SARS-CoV-1 and -2, DENV, and CHIKV, replicate in curved membrane compartments within host cells. Non-structural proteins (nsPs) critically regulate these nanoscale membrane structures, yet their curvature-dependent assembly remains elusive due to the challenges of imaging nanoscale interaction on curved surfaces. Using vertically aligned nanostructures to generate pre-defined membrane curvatures, we here investigate the impact of curvature on nsPs assembly. Taking CHIKV as a model, we reveal that nsP1 preferentially binds and stabilizes on positively curved membranes, with stronger accumulation at radii ≤150 nm. This is driven by hydrophobic residues in the membrane association (MA) loops of individual nsP1. Molecular dynamics simulations further confirm the improved binding stability of nsP1 on curved membranes, particularly when it forms a dodecamer ring. Together, nsP1 supports a strong saddle curvature association, with flexible MA loops sensing a range of positive curvatures in the x-z plane while the rigid dodecamer stabilizing fixed negative curvature in the x-y plane - crucial for constraining the membrane spherule neck during replication progression. Moreover, CHIKV replication enriches on patterned nanoring structures, underscoring the curvature-guided assembly of the viral replication complex. Our findings highlight membrane curvature as a key regulator of viral nsPs organization, opening new avenues for studying membrane remodeling in viral replication.
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spelling doaj-art-974035f9d74e46f6b4fc50ed0762db6b2025-08-20T01:49:46ZengNature PortfolioNature Communications2041-17232025-05-0116111810.1038/s41467-025-59402-0Saddle curvature association of nsP1 facilitates the replication complex assembly of Chikungunya virus in cellsXinwen Miao0Michelle Cheok Yien Law1Jatin Kumar2Choon-Peng Chng3Yongpeng Zeng4Yaw Bia Tan5Jiawei Wu6Xiangfu Guo7Lizhen Huang8Yinyin Zhuang9Weibo Gao10Changjin Huang11Dahai Luo12Wenting Zhao13School of Chemistry, Chemical Engineering and Biotechnology, Nanyang Technological UniversityLee Kong Chian School of Medicine, Nanyang Technological UniversitySchool of Mechanical and Aerospace Engineering, Nanyang Technological UniversitySchool of Mechanical and Aerospace Engineering, Nanyang Technological UniversitySchool of Chemistry, Chemical Engineering and Biotechnology, Nanyang Technological UniversityLee Kong Chian School of Medicine, Nanyang Technological UniversitySchool of Chemistry, Chemical Engineering and Biotechnology, Nanyang Technological UniversitySchool of Chemistry, Chemical Engineering and Biotechnology, Nanyang Technological UniversitySchool of Chemistry, Chemical Engineering and Biotechnology, Nanyang Technological UniversitySchool of Chemistry, Chemical Engineering and Biotechnology, Nanyang Technological UniversitySchool of Electrical & Electronic Engineering, Nanyang Technological UniversitySchool of Mechanical and Aerospace Engineering, Nanyang Technological UniversityLee Kong Chian School of Medicine, Nanyang Technological UniversitySchool of Chemistry, Chemical Engineering and Biotechnology, Nanyang Technological UniversityAbstract Positive-sense RNA viruses, including SARS-CoV-1 and -2, DENV, and CHIKV, replicate in curved membrane compartments within host cells. Non-structural proteins (nsPs) critically regulate these nanoscale membrane structures, yet their curvature-dependent assembly remains elusive due to the challenges of imaging nanoscale interaction on curved surfaces. Using vertically aligned nanostructures to generate pre-defined membrane curvatures, we here investigate the impact of curvature on nsPs assembly. Taking CHIKV as a model, we reveal that nsP1 preferentially binds and stabilizes on positively curved membranes, with stronger accumulation at radii ≤150 nm. This is driven by hydrophobic residues in the membrane association (MA) loops of individual nsP1. Molecular dynamics simulations further confirm the improved binding stability of nsP1 on curved membranes, particularly when it forms a dodecamer ring. Together, nsP1 supports a strong saddle curvature association, with flexible MA loops sensing a range of positive curvatures in the x-z plane while the rigid dodecamer stabilizing fixed negative curvature in the x-y plane - crucial for constraining the membrane spherule neck during replication progression. Moreover, CHIKV replication enriches on patterned nanoring structures, underscoring the curvature-guided assembly of the viral replication complex. Our findings highlight membrane curvature as a key regulator of viral nsPs organization, opening new avenues for studying membrane remodeling in viral replication.https://doi.org/10.1038/s41467-025-59402-0
spellingShingle Xinwen Miao
Michelle Cheok Yien Law
Jatin Kumar
Choon-Peng Chng
Yongpeng Zeng
Yaw Bia Tan
Jiawei Wu
Xiangfu Guo
Lizhen Huang
Yinyin Zhuang
Weibo Gao
Changjin Huang
Dahai Luo
Wenting Zhao
Saddle curvature association of nsP1 facilitates the replication complex assembly of Chikungunya virus in cells
Nature Communications
title Saddle curvature association of nsP1 facilitates the replication complex assembly of Chikungunya virus in cells
title_full Saddle curvature association of nsP1 facilitates the replication complex assembly of Chikungunya virus in cells
title_fullStr Saddle curvature association of nsP1 facilitates the replication complex assembly of Chikungunya virus in cells
title_full_unstemmed Saddle curvature association of nsP1 facilitates the replication complex assembly of Chikungunya virus in cells
title_short Saddle curvature association of nsP1 facilitates the replication complex assembly of Chikungunya virus in cells
title_sort saddle curvature association of nsp1 facilitates the replication complex assembly of chikungunya virus in cells
url https://doi.org/10.1038/s41467-025-59402-0
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