TY - JOUR
T1 - Electron Cryo-microscopy Structure of Ebola Virus Nucleoprotein Reveals a Mechanism for Nucleocapsid-like Assembly
AU - Su, Zhaoming
AU - Wu, Chao
AU - Shi, Liuqing
AU - Luthra, Priya
AU - Pintilie, Grigore D.
AU - Johnson, Britney
AU - Porter, Justin R.
AU - Ge, Peng
AU - Chen, Muyuan
AU - Liu, Gai
AU - Frederick, Thomas E.
AU - Binning, Jennifer M.
AU - Bowman, Gregory R.
AU - Zhou, Z. Hong
AU - Basler, Christopher F.
AU - Gross, Michael L.
AU - Leung, Daisy W.
AU - Chiu, Wah
AU - Amarasinghe, Gaya K.
N1 - Funding Information:
We thank Ms. S. Smith for general support and coordination. We also acknowledge the support of National Institutes of Health ( P41GM103832 to W.C., P41GM103422 to M.L.G., R01GM12400701 to G.R.B., R01AI123926 and U19AI109664 to C.F.B., P01AI120943 to G.K.A., and R01GM080139 to M.C. via S.J. Ludtke), the support of the Department of the Defense (Defense Threat Reduction Agency grant HDTRA1-16-1-0033 to C.F.B. [the content of the information does not necessarily reflect the position or the policy of the federal government, and no official endorsement should be inferred]), the computing resources provided by the Center for Computational and Integrative Biomedical Research of Baylor College of Medicine and the Texas Advanced Computing Center at the University of Texas at Austin ( TGMCB150009 ). G.R.B. holds a Career Award at the Scientific Interface from the Burroughs Wellcome Fund and a Packard Fellowship for Science and Engineering from the David and Lucile Packard Foundation . C.F.B. is a Georgia Research Alliance Eminent Scholar in Microbial Pathogenesis. We also acknowledge Ms. R. Roth and Drs. R. Zhang and J. Fitzpatrick for support in sample preparation and screening at the Washington University Center for Cellular Imaging supported by Washington University School of Medicine , the Children’s Discovery Institute of Washington University , and St. Louis Children’s Hospital ( CDI-CORE-2015-505 ) and the Foundation for Barnes-Jewish Hospital ( 3770 ). We also thank E.E.L. Amarasinghe and E.O.L. Amarasinghe for comments on the manuscript.
Funding Information:
We thank Ms. S. Smith for general support and coordination. We also acknowledge the support of National Institutes of Health (P41GM103832 to W.C., P41GM103422 to M.L.G., R01GM12400701 to G.R.B., R01AI123926 and U19AI109664 to C.F.B., P01AI120943 to G.K.A., and R01GM080139 to M.C. via S.J. Ludtke), the support of the Department of the Defense (Defense Threat Reduction Agency grant HDTRA1-16-1-0033 to C.F.B. [the content of the information does not necessarily reflect the position or the policy of the federal government, and no official endorsement should be inferred]), the computing resources provided by the Center for Computational and Integrative Biomedical Research of Baylor College of Medicine and the Texas Advanced Computing Center at the University of Texas at Austin (TGMCB150009). G.R.B. holds a Career Award at the Scientific Interface from the Burroughs Wellcome Fund and a Packard Fellowship for Science and Engineering from the David and Lucile Packard Foundation. C.F.B. is a Georgia Research Alliance Eminent Scholar in Microbial Pathogenesis. We also acknowledge Ms. R. Roth and Drs. R. Zhang and J. Fitzpatrick for support in sample preparation and screening at the Washington University Center for Cellular Imaging supported by Washington University School of Medicine, the Children's Discovery Institute of Washington University, and St. Louis Children's Hospital (CDI-CORE-2015-505) and the Foundation for Barnes-Jewish Hospital (3770). We also thank E.E.L. Amarasinghe and E.O.L. Amarasinghe for comments on the manuscript.
Publisher Copyright:
© 2018 Elsevier Inc.
PY - 2018/2/22
Y1 - 2018/2/22
N2 - Ebola virus nucleoprotein (eNP) assembles into higher-ordered structures that form the viral nucleocapsid (NC) and serve as the scaffold for viral RNA synthesis. However, molecular insights into the NC assembly process are lacking. Using a hybrid approach, we characterized the NC-like assembly of eNP, identified novel regulatory elements, and described how these elements impact function. We generated a three-dimensional structure of the eNP NC-like assembly at 5.8 Å using electron cryo-microscopy and identified a new regulatory role for eNP helices α22–α23. Biochemical, biophysical, and mutational analyses revealed that inter-eNP contacts within α22–α23 are critical for viral NC assembly and regulate viral RNA synthesis. These observations suggest that the N terminus and α22–α23 of eNP function as context-dependent regulatory modules (CDRMs). Our current study provides a framework for a structural mechanism for NC-like assembly and a new therapeutic target. Biochemical, biophysical, and functional validation of a 5.8 Å cryo-EM structure of the Ebola virus nucleoprotein provides insight into filovirus nucleocapsid formation.
AB - Ebola virus nucleoprotein (eNP) assembles into higher-ordered structures that form the viral nucleocapsid (NC) and serve as the scaffold for viral RNA synthesis. However, molecular insights into the NC assembly process are lacking. Using a hybrid approach, we characterized the NC-like assembly of eNP, identified novel regulatory elements, and described how these elements impact function. We generated a three-dimensional structure of the eNP NC-like assembly at 5.8 Å using electron cryo-microscopy and identified a new regulatory role for eNP helices α22–α23. Biochemical, biophysical, and mutational analyses revealed that inter-eNP contacts within α22–α23 are critical for viral NC assembly and regulate viral RNA synthesis. These observations suggest that the N terminus and α22–α23 of eNP function as context-dependent regulatory modules (CDRMs). Our current study provides a framework for a structural mechanism for NC-like assembly and a new therapeutic target. Biochemical, biophysical, and functional validation of a 5.8 Å cryo-EM structure of the Ebola virus nucleoprotein provides insight into filovirus nucleocapsid formation.
KW - Ebola virus
KW - cryo-EM
KW - nucleocapsid
KW - nucleoprotein
KW - viral RNA synthesis
UR - http://www.scopus.com/inward/record.url?scp=85042387214&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2018.02.009
DO - 10.1016/j.cell.2018.02.009
M3 - Article
C2 - 29474922
AN - SCOPUS:85042387214
SN - 0092-8674
VL - 172
SP - 966-978.e12
JO - Cell
JF - Cell
IS - 5
ER -