Atlastin-2-mediated endoplasmic reticulum membrane tethering is critical for flavivirus replication
8 affiliations
- 1Department of Microbiology and Immunology, School of Medicine, Emory University, Atlanta, Georgia, United States of America
- 2Center for Childhood Infections and Vaccines, Children’s Healthcare of Atlanta, Division of Infectious Diseases, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, United States of America
- 3Emory Vaccine Center, Emory University, Atlanta, Georgia, United States of America
- 4Leibniz Institute of Virology, Hamburg, Germany
- 5Centre Armand-Frappier Santé Biotechnologie, Institut National de la Recherche Scientifique, Laval, Quebec, Canada
- 6German Center for Infection Research, Hamburg-Borstel-Lübeck-Riems, Hamburg, Germany
- 7Centre for Structural Systems Biology (CSSB), Hamburg, Germany
- 8Faculty of Mathematics, Informatics and Natural Sciences, Department of Biology, University of Hamburg, Hamburg, Germany
Abstract
Flaviviruses (genus Orthoflavivirus) are arthropod-borne viruses which cause approximately 400 million annual global infections in humans. Flavivirus infection requires cellular machinery to facilitate replication and spread. All known flaviviruses replicate in association with the host endoplasmic reticulum (ER), where genome replication is confined within virus-induced ER invaginations called viral replication organelles (vROs). Despite the central role of these structures during flavivirus infection, the mechanisms underlying vRO biogenesis remain undefined—particularly the membrane rearrangements required for their formation. In this work, we report a conserved role for a cellular ER remodeling protein, atlastin-2 (ATL2), in the organization of vROs within infected cells. Using confocal and electron microscopy, we show that ATL2 depletion leads to a reduction in vRO spatial distribution in flavivirus-infected cells. Changes in vRO distribution corresponded with a decrease in virus production and robust induction of innate immune responses. We also demonstrate that ATL2 accumulates in areas of vRO formation during flavivirus infection. Critically, mutational analysis showed that a tethering-competent but fusion-defective ATL2 mutant was sufficient to rescue DENV and ZIKV replication in ATL2-knockout cells. Finally, targeting of ATL2 activity using synthetic peptides significantly reduced DENV replication in both immortalized and human primary cells, suggesting a possible avenue for targeting host ER functions to limit flavivirus replication. Taken together, these results show that membrane tethering plays a critical and conserved role in flavivirus infection, functioning to organize membranes for vRO biogenesis and limit cellular immune activation. Importantly, we provide evidence that ATL2-mediated membrane organization can be targeted to inhibit viral replication.
Flaviviruses reshape cell membranes to build replication sites, but how these membranes are organized remains unclear. This study shows that homotypic membrane tethering driven by the host endoplasmic reticulum protein atlastin-2 is essential for organizing these sites and supporting viral replication.
Related
- A broadly active flavivirus NS4B targeting compound that blocks NS4B-NS3 binding and replication organelle formation
- Plasma proteomics identifies an endoplasmic reticulum stress-associated molecular signature across cardiovascular-kidney-metabolic syndrome stages
- Emodin induces defensive autophagy against white spot syndrome virus infection via targeting the hemocyanin-mediated endoplasmic reticulum-mitochondrial crosstalk in shrimp
- ARID5A Is a PERK-Dependent Effector of the Endoplasmic Reticulum Stress Response That Protects Against Steatohepatitis
- Transmembrane-domain-driven heterodimer assembly licenses integrin head-domain folding in the endoplasmic reticulum
BibTeX
@article{owen2026atlastin2mediated,
title = {{Atlastin-2-mediated endoplasmic reticulum membrane tethering is critical for flavivirus replication}},
author = {Jonathan Einterz Owen and Cheyanne Lynn Bemis and Qingyi Wang and Ambarish C. Varadan and Jacob W. Vander Velden and Laura Andačić and Olus Uyar and Mansi Gupta and Christopher D. Scharer and Laurent Chatel-Chaix and Pietro Scaturro and Mehul S. Suthar and Christopher J. Neufeldt},
journal = {PLOS Biology},
year = {2026},
month = sep,
doi = {10.1371/journal.pbio.3003556},
url = {https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3003556},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Atlastin-2-mediated endoplasmic reticulum membrane tethering is critical for flavivirus replication
AU - Jonathan Einterz Owen
AU - Cheyanne Lynn Bemis
AU - Qingyi Wang
AU - Ambarish C. Varadan
AU - Jacob W. Vander Velden
AU - Laura Andačić
AU - Olus Uyar
AU - Mansi Gupta
AU - Christopher D. Scharer
AU - Laurent Chatel-Chaix
AU - Pietro Scaturro
AU - Mehul S. Suthar
AU - Christopher J. Neufeldt
PY - 2026
DA - 2026/09/15
JO - PLOS Biology
DO - 10.1371/journal.pbio.3003556
UR - https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3003556
AB - Flaviviruses (genus Orthoflavivirus) are arthropod-borne viruses which cause approximately 400 million annual global infections in humans. Flavivirus infection requires cellular machinery to facilitate replication and spread. All known flaviviruses replicate in association with the host endoplasmic reticulum (ER), where genome replication is confined within virus-induced ER invaginations called viral replication organelles (vROs). Despite the central role of these structures during flavivirus infection, the mechanisms underlying vRO biogenesis remain undefined—particularly the membrane rearrangements required for their formation. In this work, we report a conserved role for a cellular ER remodeling protein, atlastin-2 (ATL2), in the organization of vROs within infected cells. Using confocal and electron microscopy, we show that ATL2 depletion leads to a reduction in vRO spatial distribution in flavivirus-infected cells. Changes in vRO distribution corresponded with a decrease in virus production and robust induction of innate immune responses. We also demonstrate that ATL2 accumulates in areas of vRO formation during flavivirus infection. Critically, mutational analysis showed that a tethering-competent but fusion-defective ATL2 mutant was sufficient to rescue DENV and ZIKV replication in ATL2-knockout cells. Finally, targeting of ATL2 activity using synthetic peptides significantly reduced DENV replication in both immortalized and human primary cells, suggesting a possible avenue for targeting host ER functions to limit flavivirus replication. Taken together, these results show that membrane tethering plays a critical and conserved role in flavivirus infection, functioning to organize membranes for vRO biogenesis and limit cellular immune activation. Importantly, we provide evidence that ATL2-mediated membrane organization can be targeted to inhibit viral replication. Flaviviruses reshape cell membranes to build replication sites, but how these membranes are organized remains unclear. This study shows that homotypic membrane tethering driven by the host endoplasmic reticulum protein atlastin-2 is essential for organizing these sites and supporting viral replication.
SN - 1545-7885
ER -