A loop region in the N-terminal domain of ebola virus VP40 is important in viral assembly, Budding, and egress

Emmanuel Adu-Gyamfi, Smita P. Soni, Clara S. Jee, Michelle A. Digman, Enrico Gratton, Robert V. Stahelin

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Ebola virus (EBOV) causes viral hemorrhagic fever in humans and can have clinical fatality rates of ∼60%. The EBOV genome consists of negative sense RNA that encodes seven proteins including viral protein 40 (VP40). VP40 is the major Ebola virus matrix protein and regulates assembly and egress of infectious Ebola virus particles. It is well established that VP40 assembles on the inner leaflet of the plasma membrane of human cells to regulate viral budding where VP40 can produce virus like particles (VLPs) without other Ebola virus proteins present. The mechanistic details, however, of VP40 lipid-interactions and protein-protein interactions that are important for viral release remain to be elucidated. Here, we mutated a loop region in the N-terminal domain of VP40 (Lys127, Thr129, and Asn130) and find that mutations (K127A, T129A, and N130A) in this loop region reduce plasma membrane localization of VP40. Additionally, using total internal reflection fluorescence microscopy and number and brightness analysis we demonstrate these mutations greatly reduce VP40 oligomerization. Lastly, VLP assays demonstrate these mutations significantly reduce VLP release from cells. Taken together, these studies identify an important loop region in VP40 that may be essential to viral egress.

Original languageEnglish (US)
Pages (from-to)3837-3854
Number of pages18
JournalViruses
Volume6
Issue number10
DOIs
StatePublished - Oct 17 2014

Fingerprint

Ebolavirus
Virus Assembly
Viral Proteins
Virion
Proteins
Mutation
Viral Hemorrhagic Fevers
Cell Membrane
Fluorescence Microscopy
Genome
RNA
Lipids

Keywords

  • Ebola virus
  • Filovirus
  • Number and brightness analysis
  • Plasma membrane
  • Viral budding
  • Vp40

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology

Cite this

A loop region in the N-terminal domain of ebola virus VP40 is important in viral assembly, Budding, and egress. / Adu-Gyamfi, Emmanuel; Soni, Smita P.; Jee, Clara S.; Digman, Michelle A.; Gratton, Enrico; Stahelin, Robert V.

In: Viruses, Vol. 6, No. 10, 17.10.2014, p. 3837-3854.

Research output: Contribution to journalArticle

Adu-Gyamfi, Emmanuel ; Soni, Smita P. ; Jee, Clara S. ; Digman, Michelle A. ; Gratton, Enrico ; Stahelin, Robert V. / A loop region in the N-terminal domain of ebola virus VP40 is important in viral assembly, Budding, and egress. In: Viruses. 2014 ; Vol. 6, No. 10. pp. 3837-3854.
@article{78680db13f354b64b7525d0de9b37583,
title = "A loop region in the N-terminal domain of ebola virus VP40 is important in viral assembly, Budding, and egress",
abstract = "Ebola virus (EBOV) causes viral hemorrhagic fever in humans and can have clinical fatality rates of ∼60{\%}. The EBOV genome consists of negative sense RNA that encodes seven proteins including viral protein 40 (VP40). VP40 is the major Ebola virus matrix protein and regulates assembly and egress of infectious Ebola virus particles. It is well established that VP40 assembles on the inner leaflet of the plasma membrane of human cells to regulate viral budding where VP40 can produce virus like particles (VLPs) without other Ebola virus proteins present. The mechanistic details, however, of VP40 lipid-interactions and protein-protein interactions that are important for viral release remain to be elucidated. Here, we mutated a loop region in the N-terminal domain of VP40 (Lys127, Thr129, and Asn130) and find that mutations (K127A, T129A, and N130A) in this loop region reduce plasma membrane localization of VP40. Additionally, using total internal reflection fluorescence microscopy and number and brightness analysis we demonstrate these mutations greatly reduce VP40 oligomerization. Lastly, VLP assays demonstrate these mutations significantly reduce VLP release from cells. Taken together, these studies identify an important loop region in VP40 that may be essential to viral egress.",
keywords = "Ebola virus, Filovirus, Number and brightness analysis, Plasma membrane, Viral budding, Vp40",
author = "Emmanuel Adu-Gyamfi and Soni, {Smita P.} and Jee, {Clara S.} and Digman, {Michelle A.} and Enrico Gratton and Stahelin, {Robert V.}",
year = "2014",
month = "10",
day = "17",
doi = "10.3390/v6103837",
language = "English (US)",
volume = "6",
pages = "3837--3854",
journal = "Viruses",
issn = "1999-4915",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
number = "10",

}

TY - JOUR

T1 - A loop region in the N-terminal domain of ebola virus VP40 is important in viral assembly, Budding, and egress

AU - Adu-Gyamfi, Emmanuel

AU - Soni, Smita P.

AU - Jee, Clara S.

AU - Digman, Michelle A.

AU - Gratton, Enrico

AU - Stahelin, Robert V.

PY - 2014/10/17

Y1 - 2014/10/17

N2 - Ebola virus (EBOV) causes viral hemorrhagic fever in humans and can have clinical fatality rates of ∼60%. The EBOV genome consists of negative sense RNA that encodes seven proteins including viral protein 40 (VP40). VP40 is the major Ebola virus matrix protein and regulates assembly and egress of infectious Ebola virus particles. It is well established that VP40 assembles on the inner leaflet of the plasma membrane of human cells to regulate viral budding where VP40 can produce virus like particles (VLPs) without other Ebola virus proteins present. The mechanistic details, however, of VP40 lipid-interactions and protein-protein interactions that are important for viral release remain to be elucidated. Here, we mutated a loop region in the N-terminal domain of VP40 (Lys127, Thr129, and Asn130) and find that mutations (K127A, T129A, and N130A) in this loop region reduce plasma membrane localization of VP40. Additionally, using total internal reflection fluorescence microscopy and number and brightness analysis we demonstrate these mutations greatly reduce VP40 oligomerization. Lastly, VLP assays demonstrate these mutations significantly reduce VLP release from cells. Taken together, these studies identify an important loop region in VP40 that may be essential to viral egress.

AB - Ebola virus (EBOV) causes viral hemorrhagic fever in humans and can have clinical fatality rates of ∼60%. The EBOV genome consists of negative sense RNA that encodes seven proteins including viral protein 40 (VP40). VP40 is the major Ebola virus matrix protein and regulates assembly and egress of infectious Ebola virus particles. It is well established that VP40 assembles on the inner leaflet of the plasma membrane of human cells to regulate viral budding where VP40 can produce virus like particles (VLPs) without other Ebola virus proteins present. The mechanistic details, however, of VP40 lipid-interactions and protein-protein interactions that are important for viral release remain to be elucidated. Here, we mutated a loop region in the N-terminal domain of VP40 (Lys127, Thr129, and Asn130) and find that mutations (K127A, T129A, and N130A) in this loop region reduce plasma membrane localization of VP40. Additionally, using total internal reflection fluorescence microscopy and number and brightness analysis we demonstrate these mutations greatly reduce VP40 oligomerization. Lastly, VLP assays demonstrate these mutations significantly reduce VLP release from cells. Taken together, these studies identify an important loop region in VP40 that may be essential to viral egress.

KW - Ebola virus

KW - Filovirus

KW - Number and brightness analysis

KW - Plasma membrane

KW - Viral budding

KW - Vp40

UR - http://www.scopus.com/inward/record.url?scp=84908242356&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84908242356&partnerID=8YFLogxK

U2 - 10.3390/v6103837

DO - 10.3390/v6103837

M3 - Article

C2 - 25330123

AN - SCOPUS:84908242356

VL - 6

SP - 3837

EP - 3854

JO - Viruses

JF - Viruses

SN - 1999-4915

IS - 10

ER -