title: The N terminus of adenovirus type 12 E1A inhibits major histocompatibility complex class I expression by preventing phosphorylation of NF-kappaB p65 Ser276 through direct binding.
abstract: The immune-escape strategy employed by human oncogenic adenovirus type 12 (Ad12) involves downregulation of major histocompatibility complex class I (MHC-I) transcription by disabling the transactivator NF-kappaB (p50/p65). This is accomplished by the Ad12 E1A protein (E1A-12), which prevents NF-kappaB from becoming phosphorylated by the protein kinase A catalytic subunit (PKAc). In this study, we examined the interactions between E1A-12 and NF-kappaB. Our data show that an E1A-12 mutant retaining the N-terminal 66 amino acids was as effective as the wild-type E1A-12 protein (266 amino acids) in binding p65, preventing phosphorylation of p65-Ser(276), and inhibiting transactivation. In contrast, the nontumorigenic adenovirus type 5 E1A protein (E1A-5) and other E1A-12 mutants lacking the N-terminal regions were severely defective in these activities. Further studies revealed that an N-terminal peptide consisting of residues 1 to 40 of E1A-12 was able to associate directly with p65 in vitro and prevent PKAc from phosphorylating p65-Ser(276). In the absence of the N terminus, there is an almost complete loss of E1A-12 binding to p65. These findings provide solid evidence for the role of the E1A-12 N terminus as an NF-kappaB binding domain. Significantly, this study indicates that the E1A-12 N terminus prevents PKAc from gaining access to p65 to account for Ser(276) hypophosphorylation. The E1A-12 N terminus interaction with p65 serves as a key explanation of how Ad12 downregulates MHC-I transcription and contributes to oncogenesis by escaping cytotoxic T lymphocytes.
authors.affiliation: Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Junfang
authors.initials: J
authors.lastname: Jiao
authors.forename: Hancheng
authors.initials: H
authors.lastname: Guan
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Lippa
authors.forename: Robert P
authors.initials: RP
authors.lastname: Ricciardi
mesh_terms: D017500:Adenovirus E1A Proteins / Q000378:metabolism*; D000260:Adenoviruses, Human / Q000276:immunology / Q000472:pathogenicity*; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D015870:Gene Expression; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis*; D010766:Phosphorylation; D011485:Protein Binding; D025941:Protein Interaction Mapping; D051996:Transcription Factor RelA / Q000378:metabolism*; D037521:Virulence Factors / Q000378:metabolism*
chemical_list: D017500:Adenovirus E1A Proteins; D015395:Histocompatibility Antigens Class I; D051996:Transcription Factor RelA; D037521:Virulence Factors