Increased ATP generation in the host cell is required for efficient vaccinia virus production.
To search for cellular genes up-regulated by vaccinia virus (VV) infection, differential display-reverse transcription-polymerase chain reaction (ddRT-PCR) assays were used to examine the expression of mRNAs from mock-infected and VV-infected HeLa cells. Two mitochondrial genes for proteins that are part of the electron transport chain that generates ATP, ND4 and CO II, were up-regulated after VV infection. Up-regulation of ND4 level by VV infection was confirmed by Western blotting analysis. Up-regulation of ND4 was reduced by the MAPK inhibitor, apigenin, which has been demonstrated elsewhere to inhibit VV replication. The induction of ND4 expression occurred after viral DNA replication since ara C, an inhibitor of poxviral DNA replication, could block this induction. ATP production was increased in the host cells after VV infection. Moreover, 4.5 microM oligomycin, an inhibitor of ATP production, reduced the ATP level 13 hr after virus infection to that of mock-infected cells and inhibited viral protein expression and virus production, suggesting that increased ATP production is required for efficient VV production. Our results further suggest that induction of ND4 expression is through a Bcl-2 independent pathway.
Chang
Chia-Wei
CW
Graduate Institute of Molecular and Cellular Biology, Tzu Chi University, Hualien, Taiwan, Republic of China. jiawei0823@mail.tcu.edu.tw
Li
Hui-Chun
HC
Hsu
Che-Fang
CF
Chang
Chiao-Yen
CY
Lo
Shih-Yen
SY
D000255:Adenosine Triphosphate / Q000096:biosynthesis / Q000502:physiology*; D047310:Apigenin / Q000494:pharmacology; D003561:Cytarabine / Q000494:pharmacology; D004579:Electron Transport / Q000187:drug effects; D003576:Electron Transport Complex IV / Q000096:biosynthesis / Q000235:genetics; D020869:Gene Expression Profiling; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D009245:NADH Dehydrogenase / Q000096:biosynthesis / Q000235:genetics; D009840:Oligomycins / Q000494:pharmacology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000502:physiology; D034741:RNA, Small Interfering / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015290:Second Messenger Systems / Q000502:physiology*; D015854:Up-Regulation; D014616:Vaccinia virus / Q000502:physiology*; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology; D014779:Virus Replication / Q000502:physiology*
C464479:N1L protein, Vaccinia virus; C098483:NADH dehydrogenase subunit 4; D009840:Oligomycins; D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; D014764:Viral Proteins; D003561:Cytarabine; D047310:Apigenin; D000255:Adenosine Triphosphate; D009245:NADH Dehydrogenase; C117108:cytochrome C oxidase subunit II; D003576:Electron Transport Complex IV