siRNA Screen Identifies Trafficking Host Factors that Modulate Alphavirus Infection

PLoS Pathogens

Shuǐqìng Yú¹  Yíngyún Caì¹  Jens H. Kuhn¹  Robin Burk¹  Gianluca Pegoraro²  Chih-Yuan Chiang²  Xiǎolì Chī²  Jeremiah C. Clester²  David P. Langan²  Knashka Underwood²  Sina Bavari²  Krishna Kota²  Sheli R. Radoshitzky²  Christopher L. Cooper²  Rouzbeh Zamani²  Lián Dǒng²  Lucas Jozwick²  Duane Courier²  Kathleen A. Kuehl³  Mei G. Sun³ 

[1] Integrated Research Facility at Fort Detrick (IRF-Frederick), Division of Clinical Research (DCR), National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Fort Detrick, Frederick, Maryland, United States of America;Molecular and Translational Sciences Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Frederick, Maryland, United States of America;Pathology Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Frederick, Maryland, United States of America

关键词: Actins;    Small interfering RNAs;    HeLa cells;    Cell staining;    Nuclear staining;    Antibodies;    Rift Valley fever virus;    Chikungunya infection;   

DOI  :  10.1371/journal.ppat.1005466

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Little is known about the repertoire of cellular factors involved in the replication of pathogenic alphaviruses. To uncover molecular regulators of alphavirus infection, and to identify candidate drug targets, we performed a high-content imaging-based siRNA screen. We revealed an actin-remodeling pathway involving Rac1, PIP5K1- α, and Arp3, as essential for infection by pathogenic alphaviruses. Infection causes cellular actin rearrangements into large bundles of actin filaments termed actin foci. Actin foci are generated late in infection concomitantly with alphavirus envelope (E2) expression and are dependent on the activities of Rac1 and Arp3. E2 associates with actin in alphavirus-infected cells and co-localizes with Rac1–PIP5K1-α along actin filaments in the context of actin foci. Finally, Rac1, Arp3, and actin polymerization inhibitors interfere with E2 trafficking from the trans-Golgi network to the cell surface, suggesting a plausible model in which transport of E2 to the cell surface is mediated via Rac1- and Arp3-dependent actin remodeling.

【 授权许可】

CC BY