@article{8b00a49803f44ea6ae6e27f53a6cab52,
title = "Dengue NS2A Protein Orchestrates Virus Assembly",
abstract = "Dengue virus assembly requires cleavage of viral C-prM-E polyprotein into three structural proteins (capsid, premembrane, and envelope), packaging of viral RNA with C protein into nucleocapsid, and budding of prM and E proteins into virions. The molecular mechanisms underlying these assembly events are unclear. Here, we show that dengue nonstructural protein 2A (NS2A protein) recruits viral RNA, structural proteins, and protease to the site of virion assembly and coordinates nucleocapsid and virus formation. The last 285 nucleotides of viral 3′ UTR serve as a “recruiting signal for packaging” that binds to a cytosolic loop of NS2A. This interaction allows NS2A to recruit nascent RNA from the replication complex to the virion assembly site. NS2A also recruits the C-prM-E polyprotein and NS2B-NS3 protease to the virion assembly site by interacting with prM, E, and NS3, leading to coordinated C-prM-E cleavage. Mature C protein assembles onto genomic RNA to form nucleocapsid, followed by prM and E envelopment and virion formation.",
keywords = "3{\textquoteright}UTR, RNA binding, assembly, dengue virus, flavivirus, nonstructural protein 2A",
author = "Xuping Xie and Jing Zou and Xianwen Zhang and Yiyang Zhou and Routh, {Andrew L.} and Congbao Kang and Popov, {Vsevolod L.} and Xinwen Chen and Wang, {Qing Yin} and Hongping Dong and Shi, {Pei Yong}",
note = "Funding Information: We thank Drs. Mariano A. Garcia-Blanco and Shelton S. Bradrick for helpful discussions during this work. P.-Y.S. was supported by NIH grants AI142759 , AI127744 , and AI136126 and awards from the Kleberg Foundation , John S. Dunn Foundation , Amon G. Carter Foundation , Gilson Longenbaugh Foundation , and Summerfield Robert Foundation . X.X. and J.Z. were awarded with postdoctoral fellowships from the Novartis Institutes for Biomedical Research (NIBR) and the McLaughlin Endowment at University of Texas Medical Branch , respectively. Funding Information: The amino acid sequences of flavivirus NS2A proteins were download from GenBank. Sequence alignment was performed with default settings using the CLC main workbench software (Qiagen). IFA images processing and cell counting was performed in software ImageJ (National Institutes of Health, NIH). At least four different IFA images were used to estimate the mean and standard deviations of percentage of viral protein staining positive-cells in Figures 1 G and S2 . Densitometry analysis was performed using Image lab version 6.0 (Bio-Rad Laboratories, Hercules, CA). All numerical data are presented as the mean ± SD (standard deviations). Group comparisons were performed using the unpaired parametric t-test in software Prism 8.0 (GraphPad). ∗ p<0.05, significant; ∗∗ p<0.01, very significant; ∗∗∗ p<0.001, highly significant; n.s., not significant. Images were assembled using the software Adobe illustrator CC. Funding Information: We thank Drs. Mariano A. Garcia-Blanco and Shelton S. Bradrick for helpful discussions during this work. P.-Y.S. was supported by NIH grants AI142759, AI127744, and AI136126 and awards from the Kleberg Foundation, John S. Dunn Foundation, Amon G. Carter Foundation, Gilson Longenbaugh Foundation, and Summerfield Robert Foundation. X.X. and J.Z. were awarded with postdoctoral fellowships from the Novartis Institutes for Biomedical Research (NIBR) and the McLaughlin Endowment at University of Texas Medical Branch, respectively. Conceptualization, X.X. and P.-Y.S.; Methodology, X.X. J.Z. X.Z. Y.Z. A.R. V.L.P. Q.-Y.W. H.D. and C.K.; Investigation, X.X. J.Z. X.Z. Y.Z. C.K. Q.-Y.W. and H.D.; Resources, V.L.P. and C.K.; Data Curation, X.X. X.Z. Y.Z. and A.R.; Writing ? Original Draft, X.X. and P.-Y.S.; Writing ? Review & Editing, X.X. A.R. V.L.P. X.W. Q.-Y.W. H.D. and P.-Y.S.; Visualization, X.X. and P.-Y.S.; Supervision, P.-Y.S.; Funding Acquisition, P.-Y.S. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",
year = "2019",
month = nov,
day = "13",
doi = "10.1016/j.chom.2019.09.015",
language = "English (US)",
volume = "26",
pages = "606--622.e8",
journal = "Cell Host and Microbe",
issn = "1931-3128",
publisher = "Cell Press",
number = "5",
}