The ESCRT machinery is recruited by the viral BFRF1 protein to the nucleus-associated membrane for the maturation of Epstein-Barr Virus

Chung-Pei Lee; Po-Ting Liu; Hsiu-Ni Kung; Mei-Tzu Su; Huey-Huey Chua; Yu-Hsin Chang; Chou-Wei Chang; Ching-Hwa Tsai; Fu-Tong Liu; Mei-Ru Chen

doi:10.1371/journal.ppat.1002904

The ESCRT machinery is recruited by the viral BFRF1 protein to the nucleus-associated membrane for the maturation of Epstein-Barr Virus

PLoS Pathog. 2012 Sep;8(9):e1002904. doi: 10.1371/journal.ppat.1002904. Epub 2012 Sep 6.

Authors

Chung-Pei Lee¹, Po-Ting Liu, Hsiu-Ni Kung, Mei-Tzu Su, Huey-Huey Chua, Yu-Hsin Chang, Chou-Wei Chang, Ching-Hwa Tsai, Fu-Tong Liu, Mei-Ru Chen

Affiliation

¹ Graduate Institute and Department of Microbiology, College of Medicine, National Taiwan University, Taipei, Taiwan.

Abstract

The cellular endosomal sorting complex required for transport (ESCRT) machinery participates in membrane scission and cytoplasmic budding of many RNA viruses. Here, we found that expression of dominant negative ESCRT proteins caused a blockade of Epstein-Barr virus (EBV) release and retention of viral BFRF1 at the nuclear envelope. The ESCRT adaptor protein Alix was redistributed and partially colocalized with BFRF1 at the nuclear rim of virus replicating cells. Following transient transfection, BFRF1 associated with ESCRT proteins, reorganized the nuclear membrane and induced perinuclear vesicle formation. Multiple domains within BFRF1 mediated vesicle formation and Alix recruitment, whereas both Bro and PRR domains of Alix interacted with BFRF1. Inhibition of ESCRT machinery abolished BFRF1-induced vesicle formation, leading to the accumulation of viral DNA and capsid proteins in the nucleus of EBV-replicating cells. Overall, data here suggest that BFRF1 recruits the ESCRT components to modulate nuclear envelope for the nuclear egress of EBV.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Calcium-Binding Proteins / metabolism
Calcium-Binding Proteins / physiology
Cell Cycle Proteins / metabolism
Cell Cycle Proteins / physiology
Cell Line, Tumor
Cell Nucleus / metabolism*
Endosomal Sorting Complexes Required for Transport / metabolism*
Endosomal Sorting Complexes Required for Transport / physiology
Gene Expression Regulation, Viral / physiology
Herpesvirus 4, Human / genetics
Herpesvirus 4, Human / metabolism
Herpesvirus 4, Human / physiology*
Humans
Membrane Proteins / genetics
Membrane Proteins / metabolism*
Membrane Proteins / physiology
Nuclear Envelope / metabolism*
Protein Binding / genetics
Protein Transport
Tissue Distribution
Transfection
Viral Proteins / genetics
Viral Proteins / metabolism*
Viral Proteins / physiology
Virus Assembly / genetics
Virus Assembly / physiology*
Virus Release / genetics
Virus Release / physiology

Substances

BFRF1 protein, Human herpesvirus 4
Calcium-Binding Proteins
Cell Cycle Proteins
Endosomal Sorting Complexes Required for Transport
Membrane Proteins
PDCD6IP protein, human
Viral Proteins

Grants and funding

This study was supported by grants NHRI-EX99-9928BI from National Health Research Institutes (http://english.nhri.org.tw/NHRI_WEB/nhriw001Action.do), grants NSC98-2320-B002-054-MY3 and NSC100-2320-B-227-002-MY2 from National Science Council (http://web1.nsc.gov.tw/mp.aspx?mp=7) and grant 99R40044 from National Taiwan University (http://www.ntu.edu.tw/english/), Taiwan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.