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Title: Nipah Virus-Like Particle Egress Is Modulated by Cytoskeletal and Vesicular Trafficking Pathways: a Validated Particle Proteomics Analysis

Abstract

ABSTRACT Classified as a biosafety level 4 (BSL4) select agent, Nipah virus (NiV) is a deadly henipavirus in theParamyxoviridaefamily, with a nearly 75% mortality rate in humans, underscoring its global and animal health importance. Elucidating the process of viral particle production in host cells is imperative both for targeted drug design and viral particle-based vaccine development. However, little is understood concerning the functions of cellular machinery in paramyxoviral and henipaviral assembly and budding. Recent studies showed evidence for the involvement of multiple NiV proteins in viral particle formation, in contrast to the mechanisms understood for several paramyxoviruses as being reliant on the matrix (M) protein alone. Further, the levels and purposes of cellular factor incorporation into viral particles are largely unexplored for the paramyxoviruses. To better understand the involvement of cellular machinery and the major structural viral fusion (F), attachment (G), and matrix (M) proteins, we performed proteomics analyses on virus-like particles (VLPs) produced from several combinations of these NiV proteins. Our findings indicate that NiV VLPs incorporate vesicular trafficking and actin cytoskeletal factors. The involvement of these biological processes was validated by experiments indicating that the perturbation of key factors in these cellular processes substantially modulated viral particle formation.more » These effects were most impacted for NiV-F-modulated viral particle formation either autonomously or in combination with other NiV proteins, indicating that NiV-F budding relies heavily on these cellular processes. These findings indicate a significant involvement of the NiV fusion protein, vesicular trafficking, and actin cytoskeletal processes in efficient viral particle formation. IMPORTANCENipah virus is a zoonotic biosafety level 4 agent with high mortality rates in humans. The genus to which Nipah virus belongs,Henipavirus, includes five officially recognized pathogens; however, over 20 species have been identified in multiple continents within the last several years. As there are still no vaccines or treatments for NiV infection, elucidating its process of viral particle production is imperative both for targeted drug design as well as for particle-based vaccine development. Developments in high-throughput technologies make proteomic analysis of isolated viral particles a highly insightful approach to understanding the life cycle of pathogens such as Nipah virus.« less

Authors:
 [1];  [2]; ORCiD logo [3]; ORCiD logo [3];  [1];  [1];  [1];  [1];  [1]; ORCiD logo [3];  [3];  [2]; ORCiD logo [3];  [1];  [4]
  1. Cornell Univ., Ithaca, NY (United States)
  2. Washington State Univ., Pullman, WA (United States)
  3. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
  4. Harvard Medical School, Boston, MA (United States)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1591872
Alternate Identifier(s):
OSTI ID: 1591873
Report Number(s):
PNNL-SA-142825
Journal ID: ISSN 2379-5077
Grant/Contract Number:  
AC05-76RL01830; HSHQPM-14-X-00238
Resource Type:
Accepted Manuscript
Journal Name:
mSystems
Additional Journal Information:
Journal Volume: 4; Journal Issue: 5; Journal ID: ISSN 2379-5077
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Nipah virus; paramyxovirus; proteomics; vesicular trafficking; endocytosis; cytoskeleton; host-pathogen interaction

Citation Formats

Johnston, Gunner P., Bradel-Tretheway, Birgit, Piehowski, Paul D., Brewer, Heather M., Lee, Bom Nae Rin, Usher, Nicholas T., Zamora, J. Lizbeth Reyes, Ortega, Victoria, Contreras, Erik M., Teuton, Jeremy R., Wendler, Jason P., Matz, Keesha M., Adkins, Joshua N., Aguilar, Hector C., and Knipe, David M. Nipah Virus-Like Particle Egress Is Modulated by Cytoskeletal and Vesicular Trafficking Pathways: a Validated Particle Proteomics Analysis. United States: N. p., 2019. Web. https://doi.org/10.1128/mSystems.00194-19.
Johnston, Gunner P., Bradel-Tretheway, Birgit, Piehowski, Paul D., Brewer, Heather M., Lee, Bom Nae Rin, Usher, Nicholas T., Zamora, J. Lizbeth Reyes, Ortega, Victoria, Contreras, Erik M., Teuton, Jeremy R., Wendler, Jason P., Matz, Keesha M., Adkins, Joshua N., Aguilar, Hector C., & Knipe, David M. Nipah Virus-Like Particle Egress Is Modulated by Cytoskeletal and Vesicular Trafficking Pathways: a Validated Particle Proteomics Analysis. United States. https://doi.org/10.1128/mSystems.00194-19
Johnston, Gunner P., Bradel-Tretheway, Birgit, Piehowski, Paul D., Brewer, Heather M., Lee, Bom Nae Rin, Usher, Nicholas T., Zamora, J. Lizbeth Reyes, Ortega, Victoria, Contreras, Erik M., Teuton, Jeremy R., Wendler, Jason P., Matz, Keesha M., Adkins, Joshua N., Aguilar, Hector C., and Knipe, David M. Tue . "Nipah Virus-Like Particle Egress Is Modulated by Cytoskeletal and Vesicular Trafficking Pathways: a Validated Particle Proteomics Analysis". United States. https://doi.org/10.1128/mSystems.00194-19. https://www.osti.gov/servlets/purl/1591872.
@article{osti_1591872,
title = {Nipah Virus-Like Particle Egress Is Modulated by Cytoskeletal and Vesicular Trafficking Pathways: a Validated Particle Proteomics Analysis},
author = {Johnston, Gunner P. and Bradel-Tretheway, Birgit and Piehowski, Paul D. and Brewer, Heather M. and Lee, Bom Nae Rin and Usher, Nicholas T. and Zamora, J. Lizbeth Reyes and Ortega, Victoria and Contreras, Erik M. and Teuton, Jeremy R. and Wendler, Jason P. and Matz, Keesha M. and Adkins, Joshua N. and Aguilar, Hector C. and Knipe, David M.},
abstractNote = {ABSTRACT Classified as a biosafety level 4 (BSL4) select agent, Nipah virus (NiV) is a deadly henipavirus in theParamyxoviridaefamily, with a nearly 75% mortality rate in humans, underscoring its global and animal health importance. Elucidating the process of viral particle production in host cells is imperative both for targeted drug design and viral particle-based vaccine development. However, little is understood concerning the functions of cellular machinery in paramyxoviral and henipaviral assembly and budding. Recent studies showed evidence for the involvement of multiple NiV proteins in viral particle formation, in contrast to the mechanisms understood for several paramyxoviruses as being reliant on the matrix (M) protein alone. Further, the levels and purposes of cellular factor incorporation into viral particles are largely unexplored for the paramyxoviruses. To better understand the involvement of cellular machinery and the major structural viral fusion (F), attachment (G), and matrix (M) proteins, we performed proteomics analyses on virus-like particles (VLPs) produced from several combinations of these NiV proteins. Our findings indicate that NiV VLPs incorporate vesicular trafficking and actin cytoskeletal factors. The involvement of these biological processes was validated by experiments indicating that the perturbation of key factors in these cellular processes substantially modulated viral particle formation. These effects were most impacted for NiV-F-modulated viral particle formation either autonomously or in combination with other NiV proteins, indicating that NiV-F budding relies heavily on these cellular processes. These findings indicate a significant involvement of the NiV fusion protein, vesicular trafficking, and actin cytoskeletal processes in efficient viral particle formation. IMPORTANCENipah virus is a zoonotic biosafety level 4 agent with high mortality rates in humans. The genus to which Nipah virus belongs,Henipavirus, includes five officially recognized pathogens; however, over 20 species have been identified in multiple continents within the last several years. As there are still no vaccines or treatments for NiV infection, elucidating its process of viral particle production is imperative both for targeted drug design as well as for particle-based vaccine development. Developments in high-throughput technologies make proteomic analysis of isolated viral particles a highly insightful approach to understanding the life cycle of pathogens such as Nipah virus.},
doi = {10.1128/mSystems.00194-19},
journal = {mSystems},
number = 5,
volume = 4,
place = {United States},
year = {2019},
month = {9}
}

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