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Identification and characterization of SARS‐CoV protein 3a and its interacting cellular proteins
Author(s) -
Yu ChiaJung,
Wu HsuYing,
Chen JianHong,
Hsu YuTien,
Lee FangJen S.
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb101
Subject(s) - microbiology and biotechnology , golgi apparatus , hsp60 , heat shock protein , biology , immunoprecipitation , fragmentation (computing) , mitochondrion , hspa9 , transport protein , hsp70 , chemistry , biochemistry , endoplasmic reticulum , peptide sequence , gene , ecology
The severe acute respiratory syndrome coronavirus (SARS‐CoV) protein 3a is a non‐structure glycoprotein of SARS‐CoV, which with multi‐functions in host cell, including induces apoptosis, leads to cell cycle arrest at G1 phase, forms ion channel, and causes Golgi fragmentation. However, the detail molecular mechanism response for functional modulations of protein 3a remains unknown. Herein, we found that expression of non‐tagged protein 3a in Vero E6 cells reduced the mitochondria membrane potential, caused Golgi fragmentation, and impaired the vesicle transport. Using in vitro pull‐down assay combined with mass spectrometry, we identified 23 potential protein 3a‐interacting cellular proteins. Based on biological classification, we selected 60 kDa heat shock protein (Hsp60) and SNAPs association protein (SAP) for further characterization. Biochemical fractionation indicated that Hsp60 protein level in mitochondria fraction was increased in protein 3a‐expressing cells. Overexpressed SAP was partially co‐localized with protein 3a and co‐immunoprecipitated with protein 3a. Collectively, our results implied that the effect of protein 3a on the function of mitochondria and the Golgi might through its interaction with Hsp60 and SAP, respectively.