Inhibition of interferon-inducible gene expression by adenovirus E1A proteins: block in transcriptional complex formation.
Infection with wild-type adenovirus 5, but not with a mutant lacking the E1A gene, prevented the induction by interferon (IFN) alpha of chloramphenicol acetyltransferase (CAT) activity in HeLaM cell lines that had been permanently transfected with chimeric CAT reporter genes driven by the transcriptional regulatory regions of the IFN-responsive genes 561 and 6-16. Similar inhibition of IFN-inducible CAT activity was observed in cells that were cotransfected with the same reporter genes and plasmids expressing either the E1A 289- or 243-amino acid protein. These proteins also prevented the induction of CAT activity by IFN-gamma from a cotransfected HLA-DR alpha-CAT gene. Experiments with E1A mutants mapped the inhibitory activity to amino acid residues 38-65 of these proteins. In a HeLa cell line permanently expressing the E1A 289-amino acid protein, the replication of vesicular stomatitis virus and encephalomyocarditis virus was not inhibited by IFN-alpha, suggesting a global blockade of IFN responses. In accord with this theory, induction of 561, 1-8, and (2'-5')oligoadenylate synthetase mRNAs by IFN was blocked in these cells at the transcriptional level. The observed transcriptional inhibition could be attributed to the lack of formation of the crucial IFN-stimulated gene factor 3 (ISGF3) transcriptional complex. As shown by mobility shift assays, this complex was not formed in the nuclear extracts of IFN-treated adenovirus-infected cells or IFN-treated E1A-producing cells. These nuclear extracts were deficient in both ISGF3 alpha and ISGF3 gamma subunits. However, they did not block the formation of ISGF3 complex from exogenously added components.
Kalvakolanu
D V
DV
Department of Molecular Biology, Cleveland Clinic Foundation, OH 44195.
Bandyopadhyay
S K
SK
Harter
M L
ML
Sen
G C
GC
D017456:Adenovirus Early Proteins; D000260:Adenoviruses, Human / Q000235:genetics* / Q000378:metabolism; D015152:Blotting, Northern; D002472:Cell Transformation, Viral; D002678:Chimera; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors; D006367:HeLa Cells / Q000502:physiology; D006801:Humans; D007370:Interferon Type I / Q000494:pharmacology*; D007700:Kinetics; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication / Q000187:drug effects
D017456:Adenovirus Early Proteins; D007370:Interferon Type I; D009856:Oncogene Proteins, Viral; D014157:Transcription Factors; D015500:Chloramphenicol O-Acetyltransferase