HSV-1 US3 Hijacks Conserved Actin Regulatory Complexes To Drive F-Actin Remodeling

Author

• Md Imran Hossain
• Md Arifuzzaman, Missouri University of Science and TechnologyFollow
• Md Mehedi Hasan
• Seung Jong Park, Missouri University of Science and TechnologyFollow
• Leila Rahimian
• Ojasvi Dutta
• Vladimir Chouljenko
• Harikrishnan Mohan
• Reza Ghavimi
• Konstantin G. Kousoulas

Abstract

The herpes simplex virus 1 (HSV-1) US3 is a multifunctional serine/threonine kinase that promotes HSV-1 replication and spread. But its role and the mechanisms by which US3 regulates actin cytoskeletal remodeling remain poorly defined. We combined flow cytometry, confocal microscopy, immunoprecipitation-mass spectrometry (IP-MS), protein complex mapping, and machine learning to characterize US3-mediated F-actin dynamics. Flow cytometry and confocal microscopy showed that wild-type HSV-1 induces significant F-actin remodeling, while the ΔUS3 mutant displays F-actin levels comparable to uninfected cells, identifying US3 as a key regulator. IP-MS identified 47 high-confidence US3 interactors enriched in conserved actin regulatory complexes, including Arp2/3 nucleation machinery, formin-associated assemblies, cofilin severing complexes, and Rho-family GTPase modules. Mapping interactors to the CORUM database revealed clustering within actin nucleation, polymerization, and severing complexes, indicating that US3 operates through organized cellular machines. Machine-learning classifiers trained on label-independent mass-spectrometry features were used to prioritize interactors resembling known actin regulators; under protein-group-aware cross-validation, logistic regression performed best (average precision 0.24; ROC-AUC 0.66), and the analysis was interpreted as prioritization rather than de novo discovery. Pharmacological inhibition of Arp2/3 and formin pathways significantly attenuated US3-dependent F-actin remodeling, supporting the functional involvement of these pathways. Together, these findings are consistent with an inferred hierarchical axis in which US3 modulates Rho GTPase signaling and cofilin activation to promote F-actin disassembly, coordinating cytoskeletal remodeling required for efficient viral egress and spread.

Recommended Citation

M. I. Hossain and M. Arifuzzaman and M. M. Hasan and S. J. Park and L. Rahimian and O. Dutta and V. Chouljenko and H. Mohan and R. Ghavimi and K. G. Kousoulas, "HSV-1 US3 Hijacks Conserved Actin Regulatory Complexes To Drive F-Actin Remodeling," Viruses, vol. 18, no. 7, article no. 793, MDPI, Jul 2026.

The definitive version is available at https://doi.org/10.3390/v18070793

Department(s)

Computer Science

Publication Status

Open Access

Comments

Louisiana Board of Regents, Grant P20GM103424

Keywords and Phrases

CORUM; cytoskeleton regulation; F-actin remodeling; herpes simplex virus; machine learning; protein complexes; US3 kinase

International Standard Serial Number (ISSN)

1999-4915

Document Type

Article - Journal

Document Version

Final Version

File Type

text

Language(s)

English

Rights

© 2026 The Authors, All rights reserved.

Creative Commons Licensing

This work is licensed under a Creative Commons Attribution 4.0 License.

Publication Date

01 Jul 2026

PubMed ID

42515645