TY - JOUR
T1 - Open reading frame 8a of the human severe acute respiratory syndrome coronavirus not only promotes viral replication but also induces apoptosis
AU - Chen, Chia Yen
AU - Ping, Yueh Hsin
AU - Lee, Hsin Chen
AU - Chen, Kuan Hsuan
AU - Lee, Yuan Ming
AU - Chan, Yu Juin
AU - Lien, Te Cheng
AU - Jap, Tjin Shing
AU - Lin, Chi Hung
AU - Kao, Lung Sen
AU - Chen, Yi Ming Arthur
N1 - Funding Information:
Received 28 September 2006; accepted 23 January 2007; electronically published 19 June 2007. Potential conflicts of interest: none reported. Presented in part: Fourteenth Symposium on the Recent Advances in Cellular and Molecular Biology, Kenting, Taiwan, 18–20 January 2006 (poster 204). Financial support: Veterans General Hospitals University System of Taiwan Joint Research Program, Tsou Foundation (grant VGHUST95-P7-22). Reprints or correspondence: Prof. Yi-Ming A Chen, AIDS Prevention and Research Center, National Yang-Ming University, Li-Noun Street, Section 2, Taipei, Taiwan 112 ([email protected]).
PY - 2007/8/1
Y1 - 2007/8/1
N2 - Background. A unique genomic difference between human and civet severe acute respiratory syndrome coronaviruses (SARS-CoVs) is that the former has a deletion of 29 nucleotides from open reading frame (orf) 8d that results in the generation of orf8a and orf8b. The objectives of the present study were to analyze antibody reactivity to ORF8a in patients with SARS and to elucidate the function of ORF8a. Methods. Western-blot and immunofluorescent antibody assays were used to detect anti-ORF8a antibody. SARS-CoV HKU39849 was used to infect stable clones expressing ORF8a and cells transfected with small interfering RNA (siRNA). The virus loads (VLs) and cytopathic effects (CPEs) were recorded. Confocal microscopy and several mitochondria-related tests were used to study the function of ORF8a. Results. Two (5.4%) of 37 patients with SARS had anti-ORF8a antibodies. The VLs in the stable clones expressing ORF8a were significantly higher than those in control subjects 5 days after infection. siRNA against orf8a significantly reduced VLs and interrupted the CPE. ORF8a was found to be localized in mitochondria, and overexpression resulted in increases in mitochondrial transmembrane potential, reactive oxygen species production, caspase 3 activity, and cellular apoptosis. Conclusions. ORF8a not only enhances viral replication but also induces apoptosis through a mitochondria-dependent pathway.
AB - Background. A unique genomic difference between human and civet severe acute respiratory syndrome coronaviruses (SARS-CoVs) is that the former has a deletion of 29 nucleotides from open reading frame (orf) 8d that results in the generation of orf8a and orf8b. The objectives of the present study were to analyze antibody reactivity to ORF8a in patients with SARS and to elucidate the function of ORF8a. Methods. Western-blot and immunofluorescent antibody assays were used to detect anti-ORF8a antibody. SARS-CoV HKU39849 was used to infect stable clones expressing ORF8a and cells transfected with small interfering RNA (siRNA). The virus loads (VLs) and cytopathic effects (CPEs) were recorded. Confocal microscopy and several mitochondria-related tests were used to study the function of ORF8a. Results. Two (5.4%) of 37 patients with SARS had anti-ORF8a antibodies. The VLs in the stable clones expressing ORF8a were significantly higher than those in control subjects 5 days after infection. siRNA against orf8a significantly reduced VLs and interrupted the CPE. ORF8a was found to be localized in mitochondria, and overexpression resulted in increases in mitochondrial transmembrane potential, reactive oxygen species production, caspase 3 activity, and cellular apoptosis. Conclusions. ORF8a not only enhances viral replication but also induces apoptosis through a mitochondria-dependent pathway.
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U2 - 10.1086/519166
DO - 10.1086/519166
M3 - Article
C2 - 17597455
AN - SCOPUS:34447620646
SN - 0022-1899
VL - 196
SP - 405
EP - 415
JO - Journal of Infectious Diseases
JF - Journal of Infectious Diseases
IS - 3
ER -