Significance

ZIKV has “silently” circulated without causing severe diseases for decades since its discovery in 1947. Our study demonstrated that ZIKV acquired an evolutionary mutation in viral envelope gene (E-V473M) that increases virulence, maternal-to-fetal transmission during pregnancy, and viremia in nonhuman primates to facilitate urban transmission since 2013, which may be responsible for the recent emergence and severe diseases. Our results underscore the potential that high genetic mutation frequencies during arbovirus replication and transmission between mosquito and vertebrate hosts could lead to emergence and reemergence of those pathogens. Understanding the mechanisms of emergence and enhanced transmission is essential to detect and respond to future arbovirus outbreaks.

Abstract

Arboviruses maintain high mutation rates due to lack of proofreading ability of their viral polymerases, in some cases facilitating adaptive evolution and emergence. Here we show that, just before its 2013 spread to the Americas, Zika virus (ZIKV) underwent an envelope protein V473M substitution (E-V473M) that increased neurovirulence, maternal-to-fetal transmission, and viremia to facilitate urban transmission. A preepidemic Asian ZIKV strain (FSS13025 isolated in Cambodia in 2010) engineered with the V473M substitution significantly increased neurovirulence in neonatal mice and produced higher viral loads in the placenta and fetal heads in pregnant mice. Conversely, an epidemic ZIKV strain (PRVABC59 isolated in Puerto Rico in 2015) engineered with the inverse M473V substitution reversed the pathogenic phenotypes. Although E-V473M did not affect oral infection of Aedes aegypti mosquitoes, competition experiments in cynomolgus macaques showed that this mutation increased its fitness for viremia generation, suggesting adaptive evolution for human viremia and hence transmission. Mechanistically, the V473M mutation, located at the second transmembrane helix of the E protein, enhances virion morphogenesis. Overall, our study revealed E-V473M as a critical determinant for enhanced ZIKV virulence, intrauterine transmission during pregnancy, and viremia to facilitate urban transmission.

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Data Availability

Source data for generating main figures are available in the SI Appendix.

Acknowledgments

We thank Michael S. Diamond, Gong Cheng, and Cheng-Feng Qin for helpful discussions and support during the course of the work. P.-Y.S. was supported by NIH Grants AI142759, AI134907, AI145617, and UL1TR001439 and awards from the Sealy & Smith Foundation, Kleberg Foundation, John S. Dunn Foundation, Amon G. Carter Foundation, Gillson Longenbaugh Foundation, and Summerfield Robert Foundation. This research was also supported by NIH Grant AI120942 and the Western Gulf Center of Excellence for Vector-Borne Diseases (Centers for Disease Control and Prevention Grant U01CK000512) to S.C.W.

Supporting Information

Appendix (PDF)

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Information & Authors

Information

Published in

Go to Proceedings of the National Academy of Sciences
Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 117 | No. 33
August 18, 2020
PubMed: 32747564

Classifications

Data Availability

Source data for generating main figures are available in the SI Appendix.

Submission history

Published online: August 3, 2020
Published in issue: August 18, 2020

Keywords

  1. Zika
  2. envelope
  3. evolution
  4. transmission

Acknowledgments

We thank Michael S. Diamond, Gong Cheng, and Cheng-Feng Qin for helpful discussions and support during the course of the work. P.-Y.S. was supported by NIH Grants AI142759, AI134907, AI145617, and UL1TR001439 and awards from the Sealy & Smith Foundation, Kleberg Foundation, John S. Dunn Foundation, Amon G. Carter Foundation, Gillson Longenbaugh Foundation, and Summerfield Robert Foundation. This research was also supported by NIH Grant AI120942 and the Western Gulf Center of Excellence for Vector-Borne Diseases (Centers for Disease Control and Prevention Grant U01CK000512) to S.C.W.

Notes

This article is a PNAS Direct Submission.

Authors

Affiliations

Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555;
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 430071 Wuhan, China;
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555;
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Translational Science, University of Texas Medical Branch, Galveston, TX 77555;
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Translational Science, University of Texas Medical Branch, Galveston, TX 77555;
Yang Liu
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555;
Jianying Liu
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX 77555;
Antonio E. Muruato
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555;
Rubing Chen
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Translational Science, University of Texas Medical Branch, Galveston, TX 77555;
Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555;
Shannan L. Rossi
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX 77555;
Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555;
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX 77555;
Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555;
World Reference Center of Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX 77555;
Center for Biodefence and Emerging Infectious Diseases, University of Texas Medical Branch, Galveston, TX 77555;
Center for Tropical Diseases, University of Texas Medical Branch, Galveston, TX 77555;
Rongjuan Pei
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 430071 Wuhan, China;
Camila R. Fontes-Garfias
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555;
Sanjay Kumar Singh
Department of Neurosurgery-Research, The University of Texas MD Anderson Cancer Center, Houston, TX 77030;
Xuping Xie
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555;
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Translational Science, University of Texas Medical Branch, Galveston, TX 77555;
Sealy Institute for Vaccine Sciences, University of Texas Medical Branch, Galveston, TX 77555;
Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, TX 77555
Pei-Yong Shi2 [email protected]
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX 77555;
Institute for Translational Science, University of Texas Medical Branch, Galveston, TX 77555;
Sealy Institute for Vaccine Sciences, University of Texas Medical Branch, Galveston, TX 77555;
Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, TX 77555

Notes

2
To whom correspondence may be addressed. Email: [email protected], [email protected], or [email protected].
Author contributions: C.S., H.X., S.L.H., S.R.A., Y.L., S.L.R., N.V., X.X., S.C.W., and P.-Y.S. designed research; C.S., H.X., S.L.H., S.R.A., Y.L., J.L., A.E.M., R.C., S.L.R., M.W., N.V., R.P., C.R.F.-G., S.K.S., and X.X. performed research; C.S., H.X., S.L.H., S.R.A., Y.L., J.L., A.E.M., R.C., S.L.R., M.W., N.V., R.P., C.R.F.-G., S.K.S., X.X., S.C.W., and P.-Y.S. analyzed data; and C.S., H.X., S.L.H., S.R.A., S.C.W., and P.-Y.S. wrote the paper.
1
C.S. and H.X. contributed equally to this work.

Competing Interests

The authors declare no competing interest.

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    A Zika virus envelope mutation preceding the 2015 epidemic enhances virulence and fitness for transmission
    Proceedings of the National Academy of Sciences
    • Vol. 117
    • No. 33
    • pp. 19609-20337

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