title: The Cellular Protein Complex Associated with a Transforming Region of E1A Contains c-MYC.
abstract: The cell-transforming activity of human adenovirus 5 (hAd5) E1A is mediated by the N-terminal half of E1A, which interacts with three different major cellular protein complexes, p300/CBP, TRRAP/p400, and pRb family members. Among these protein interactions, the interaction of pRb family proteins with conserved region 2 (CR2) of E1A is known to promote cell proliferation by deregulating the activities of E2F family transcription factors. The functional consequences of interaction with the other two protein complexes in regulating the transforming activity of E1A are not well defined. Here, we report that the E1A N-terminal region also interacted with the cellular proto-oncoprotein c-MYC and the homolog of enhancer of yellow 2 (ENY2). Our results suggested that these proteins interacted with an essential E1A transforming domain spanning amino acid residues 26 to 35 which also interacted with TRRAP and p400. Small interfering RNA (siRNA)-mediated depletion of TRRAP reduced c-MYC interaction with E1A, while p400 depletion did not. In contrast, depletion of TRRAP enhanced ENY2 interaction with E1A, suggesting that ENY2 and TRRAP may interact with E1A in a competitive manner. The same E1A region additionally interacted with the constituents of a deubiquitinase complex consisting of USP22, ATXN7, and ATXN7L3 via TRRAP. Acute short hairpin RNA (shRNA)-mediated depletion of c-MYC reduced the E1A transforming activity, while depletion of ENY2 and MAX did not. These results suggested that the association of c-MYC with E1A may, at least partially, play a role in the E1A transformation activity, independently of MAX.
  
  
  OBJECTIVE
  The transforming region of adenovirus E1A consists of three short modules which complex with different cellular protein complexes. The mechanism by which one of the transforming modules, CR2, promotes cell proliferation, through inactivating the activities of the pRb family proteins, is better understood than the activities of the other domains. Our analysis of the E1A proteome revealed the presence of the proto-oncoprotein c-MYC and of ENY2. We mapped these interactions to a critical transforming module of E1A that was previously known to interact with the scaffolding molecule TRRAP and the E1A-binding protein p400. We showed that c-MYC interacted with E1A through TRRAP, while ENY2 interacted with it independently. The data reported here indicated that depletion of c-MYC in normal human cells reduced the transforming activity of E1A. Our result raises a novel paradigm in oncogenic transformation by a DNA viral oncogene, the E1A gene, that may exploit the activity of a cellular oncogene, the c-MYC gene, in addition to inactivation of the tumor suppressors, such as pRb.
authors.affiliation: Institute for Molecular Virology, Department of Molecular Microbiology & Immunology, Doisy Research Center, Saint Louis University School of Medicine, St. Louis, Missouri, USA.
authors.forename: S
authors.initials: S
authors.lastname: Vijayalingam
authors.affiliation: Institute for Molecular Virology, Department of Molecular Microbiology & Immunology, Doisy Research Center, Saint Louis University School of Medicine, St. Louis, Missouri, USA.
authors.forename: T
authors.initials: T
authors.lastname: Subramanian
authors.affiliation: Institute for Molecular Virology, Department of Molecular Microbiology & Immunology, Doisy Research Center, Saint Louis University School of Medicine, St. Louis, Missouri, USA.
authors.forename: Ling-Jun
authors.initials: LJ
authors.lastname: Zhao
authors.affiliation: Institute for Molecular Virology, Department of Molecular Microbiology & Immunology, Doisy Research Center, Saint Louis University School of Medicine, St. Louis, Missouri, USA chinnag@slu.edu.
authors.forename: G
authors.initials: G
authors.lastname: Chinnadurai
mesh_terms: D017500:Adenovirus E1A Proteins / Q000378:metabolism*; D000260:Adenoviruses, Human / Q000502:physiology*; D002460:Cell Line; D054884:Host-Pathogen Interactions; D006801:Humans; D011485:Protein Binding; D025941:Protein Interaction Mapping; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D017500:Adenovirus E1A Proteins; C527171:Eny2 protein, human; C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; D014157:Transcription Factors