Flavivirus NS4B proteins do not form homodimers: discrepancies with an AlphaFold-based oligomeric model

In flaviviruses such as Dengue, Zika or West Nile virus (DENV, ZIKV or WNV), non-structural protein 4B (NS4B) participates in membrane remodeling during infection and is critical for virulence and host immunity. Despite the important roles and confirmed drug target status for NS4B, no detailed struc...

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Main Authors: Wahyu Surya, Paula Tan, Shwe Sin Honey, Devanshu Mehta, Jaume Torres
Format: Article
Language:English
Published: Elsevier 2025-01-01
Series:Computational and Structural Biotechnology Journal
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Online Access:http://www.sciencedirect.com/science/article/pii/S2001037025001497
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author Wahyu Surya
Paula Tan
Shwe Sin Honey
Devanshu Mehta
Jaume Torres
author_facet Wahyu Surya
Paula Tan
Shwe Sin Honey
Devanshu Mehta
Jaume Torres
author_sort Wahyu Surya
collection DOAJ
description In flaviviruses such as Dengue, Zika or West Nile virus (DENV, ZIKV or WNV), non-structural protein 4B (NS4B) participates in membrane remodeling during infection and is critical for virulence and host immunity. Despite the important roles and confirmed drug target status for NS4B, no detailed structure is available. Flavivirus NS4B proteins have five hydrophobic domains and have been proposed to form homodimers. Herein, we have used AlphaFold to explore its multimeric organization in a diverse set of five sequences of flaviviruses. AlphaFold correctly predicts the α-helical segments identified by solution NMR, but with a fold that differs from current models involving regular transmembrane helices. Oligomers of increasing size were evaluated using a combined AlphaFold score, with optimal value centered around decamers (n = 10). We tested this model experimentally using ZIKV NS4B protein. Biochemical and biophysical data produced an oligomer larger than the current dimeric model, at least pentameric, and possibly hexameric. Overall, our results suggest a ‘regular’ NS4B oligomer in ZIKV that is at least pentameric with five regular transmembrane helices. Although no evidence was found for the ‘irregular’ Alphafold-predicted model, it is intriguing that it is common to all sequences tested. Interference with the formation of an NS4B oligomer may be a general novel approach to treat flavivirus infection.
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spelling doaj-art-410f5fa2107a4d44a2569c6bcf3ea9b02025-08-20T03:13:59ZengElsevierComputational and Structural Biotechnology Journal2001-03702025-01-01271660167210.1016/j.csbj.2025.04.026Flavivirus NS4B proteins do not form homodimers: discrepancies with an AlphaFold-based oligomeric modelWahyu Surya0Paula Tan1Shwe Sin Honey2Devanshu Mehta3Jaume Torres4School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, SingaporeSchool of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, SingaporeSchool of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, SingaporeSchool of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, SingaporeCorresponding author.; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, SingaporeIn flaviviruses such as Dengue, Zika or West Nile virus (DENV, ZIKV or WNV), non-structural protein 4B (NS4B) participates in membrane remodeling during infection and is critical for virulence and host immunity. Despite the important roles and confirmed drug target status for NS4B, no detailed structure is available. Flavivirus NS4B proteins have five hydrophobic domains and have been proposed to form homodimers. Herein, we have used AlphaFold to explore its multimeric organization in a diverse set of five sequences of flaviviruses. AlphaFold correctly predicts the α-helical segments identified by solution NMR, but with a fold that differs from current models involving regular transmembrane helices. Oligomers of increasing size were evaluated using a combined AlphaFold score, with optimal value centered around decamers (n = 10). We tested this model experimentally using ZIKV NS4B protein. Biochemical and biophysical data produced an oligomer larger than the current dimeric model, at least pentameric, and possibly hexameric. Overall, our results suggest a ‘regular’ NS4B oligomer in ZIKV that is at least pentameric with five regular transmembrane helices. Although no evidence was found for the ‘irregular’ Alphafold-predicted model, it is intriguing that it is common to all sequences tested. Interference with the formation of an NS4B oligomer may be a general novel approach to treat flavivirus infection.http://www.sciencedirect.com/science/article/pii/S2001037025001497FlavivirusDengueZikaNon-structural protein 4BOligomerizationAlphaFold
spellingShingle Wahyu Surya
Paula Tan
Shwe Sin Honey
Devanshu Mehta
Jaume Torres
Flavivirus NS4B proteins do not form homodimers: discrepancies with an AlphaFold-based oligomeric model
Computational and Structural Biotechnology Journal
Flavivirus
Dengue
Zika
Non-structural protein 4B
Oligomerization
AlphaFold
title Flavivirus NS4B proteins do not form homodimers: discrepancies with an AlphaFold-based oligomeric model
title_full Flavivirus NS4B proteins do not form homodimers: discrepancies with an AlphaFold-based oligomeric model
title_fullStr Flavivirus NS4B proteins do not form homodimers: discrepancies with an AlphaFold-based oligomeric model
title_full_unstemmed Flavivirus NS4B proteins do not form homodimers: discrepancies with an AlphaFold-based oligomeric model
title_short Flavivirus NS4B proteins do not form homodimers: discrepancies with an AlphaFold-based oligomeric model
title_sort flavivirus ns4b proteins do not form homodimers discrepancies with an alphafold based oligomeric model
topic Flavivirus
Dengue
Zika
Non-structural protein 4B
Oligomerization
AlphaFold
url http://www.sciencedirect.com/science/article/pii/S2001037025001497
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