title: Normal human cell proteins that interact with the adenovirus type 5 E1B 55kDa protein.
abstract: Several of the functions of the human adenovirus type 5 E1B 55kDa protein are fulfilled via the virus-specific E3 ubiquitin ligase it forms with the viral E4 Orf6 protein and several cellular proteins. Important substrates of this enzyme have not been identified, and other functions, including repression of transcription of interferon-sensitive genes, do not require the ligase. We therefore used immunoaffinity purification and liquid chromatography-mass spectrometry of lysates of normal human cells infected in parallel with HAdV-C5 and E1B 55kDa protein-null mutant viruses to identify specifically E1B 55kDa-associated proteins. The resulting set of >90 E1B-associated proteins contained the great majority identified previously, and was enriched for those associated with the ubiquitin-proteasome system, RNA metabolism and the cell cycle. We also report very severe inhibition of viral genome replication when cells were exposed to both specific or non-specific siRNAs and interferon prior to infection.
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA.
authors.forename: George
authors.initials: G
authors.lastname: Hung
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA. Electronic address: sjflint@princeton.edu.
authors.forename: S J
authors.initials: SJ
authors.lastname: Flint
mesh_terms: D000072283:A549 Cells; D017501:Adenovirus E1B Proteins / Q000235:genetics / Q000378:metabolism*; D017504:Adenovirus E4 Proteins / Q000378:metabolism*; D000260:Adenoviruses, Human / Q000235:genetics*; D000595:Amino Acid Sequence / Q000235:genetics; D004279:DNA, Viral / Q000096:biosynthesis; D016679:Genome, Viral / Q000235:genetics; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D009687:Nuclear Proteins; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D016601:RNA-Binding Proteins; D012097:Repressor Proteins / Q000235:genetics; D056264:Sin3 Histone Deacetylase and Corepressor Complex; D014779:Virus Replication / Q000235:genetics*
chemical_list: C087425:ANP32A protein, human; D017501:Adenovirus E1B Proteins; D017504:Adenovirus E4 Proteins; D004279:DNA, Viral; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D012097:Repressor Proteins; C099683:SIN3A transcription factor; D056264:Sin3 Histone Deacetylase and Corepressor Complex
keywords: E1B 55kDa protein; Human adenovirus type 5 (HAdV-C5); Mass spectrometry; Normal human cells; RNA metabolism; Type I interferons; Ubiquitin-proteasome system