Single molecule super resolution microscopy reveals formation of Dengue NS2B3 cluster on mitochondrial network and its effect on fragmentation
- 1Department of Instrumentation and Applied Physics, Indian Institute of Science
Abstract
It is challenging to understand viral proteins at a single-molecule level and visualize their dynamics in a cellular environment. This calls for sophisticated microscopy techniques (such as,SMLM) and single-molecule analysis that can reveal details with the precision of a single molecule. In Dengue type-2(D e n v − 2) infection, NS2B3 is recognized as a critical protein complex responsible for proteolytic activity and processing of viral polyprotein. We study the distribution of NS2B3 protein and its clusters. Accordingly, two distinct photoactivatable probes(m E o s − N S2B3 and P A G F P − N S2B3) and a fluorescent probe (eGFP-NS2B3) were constructed. Results confirmed the formation of NS2B3 clusters in close vicinity to the mitochondrial network. Statistical analysis of single-molecule data (images) facilitated the estimation of critical cluster parameters(such as cluster density, number of molecules/cluster,and its spread). In addition, regional analysis suggests a correlation between NS2B3 cluster formation and fragmentation of mitochondrial network which is further supported by time-lapse confocal imaging. In other words, the presence of cluster depletes mitochondrial segment. To further demonstrate, time-lapse super-resolution imaging is carried out that enabled visualization of the formation of NS2B3 clusters. Understanding the underlying dynamics of NS2B3 clustering at the single-molecule level may facilitate the identification of potential drug targets,and methods to disrupt them.
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BibTeX
@article{varghese2026single,
title = {{Single molecule super resolution microscopy reveals formation of Dengue NS2B3 cluster on mitochondrial network and its effect on fragmentation}},
author = {Jiby M. Varghese and Prakash Joshi and Aravinth S and Partha Pratim Mondal},
journal = {npj Viruses},
year = {2026},
month = jun,
doi = {10.1038/s44298-026-00202-8},
url = {https://doi.org/10.1038/s44298-026-00202-8},
note = {CC BY},
}
RIS
TY - JOUR
TI - Single molecule super resolution microscopy reveals formation of Dengue NS2B3 cluster on mitochondrial network and its effect on fragmentation
AU - Jiby M. Varghese
AU - Prakash Joshi
AU - Aravinth S
AU - Partha Pratim Mondal
PY - 2026
DA - 2026/06/01
JO - npj Viruses
DO - 10.1038/s44298-026-00202-8
UR - https://doi.org/10.1038/s44298-026-00202-8
AB - It is challenging to understand viral proteins at a single-molecule level and visualize their dynamics in a cellular environment. This calls for sophisticated microscopy techniques (such as,SMLM) and single-molecule analysis that can reveal details with the precision of a single molecule. In Dengue type-2(D e n v − 2) infection, NS2B3 is recognized as a critical protein complex responsible for proteolytic activity and processing of viral polyprotein. We study the distribution of NS2B3 protein and its clusters. Accordingly, two distinct photoactivatable probes(m E o s − N S2B3 and P A G F P − N S2B3) and a fluorescent probe (eGFP-NS2B3) were constructed. Results confirmed the formation of NS2B3 clusters in close vicinity to the mitochondrial network. Statistical analysis of single-molecule data (images) facilitated the estimation of critical cluster parameters(such as cluster density, number of molecules/cluster,and its spread). In addition, regional analysis suggests a correlation between NS2B3 cluster formation and fragmentation of mitochondrial network which is further supported by time-lapse confocal imaging. In other words, the presence of cluster depletes mitochondrial segment. To further demonstrate, time-lapse super-resolution imaging is carried out that enabled visualization of the formation of NS2B3 clusters. Understanding the underlying dynamics of NS2B3 clustering at the single-molecule level may facilitate the identification of potential drug targets,and methods to disrupt them.
SN - 2948-1767
ER -