title: Identification of integrin alpha3 as a new substrate of the adenovirus E4orf6/E1B 55-kilodalton E3 ubiquitin ligase complex.
abstract: The human adenovirus E4orf6 and E1B55K proteins promote viral replication by targeting several cellular proteins for degradation. The E4orf6 product has been shown by our group and others to form an E3 ubiquitin ligase complex that contains elongins B and C and cullin family member Cul5. E1B55K associates with this complex, where it is believed to function primarily to introduce bound substrates for degradation via proteasomes. In addition to p53, its first known substrate, the E4orf6/E1B 55-kDa complex (E4orf6/E1B55K) was shown to promote the degradation of Mre11 and DNA ligase IV; however, additional substrates are believed to exist. This notion is strengthened by the fact that none of these substrates seems likely to be associated with additional functions shown to be mediated by the E4orf6-associated E3 ubiquitin ligase complex, including export of late viral mRNAs and blockage of export of the bulk cellular mRNAs from the nucleus. In an attempt to identify new E4orf6/E1B55K substrates, we undertook a proteomic screen using human p53-null, non-small-cell lung carcinoma H1299 cells expressing either E4orf6 protein alone or in combination with E1B55K through infection by appropriate adenovirus vectors. One cellular protein that appeared to be degraded by E1B55K in combination with the E4orf6 protein was a species of molecular mass approximately 130 kDa that was identified as the integrin alpha3 subunit (i.e., very late activation antigen 3 alpha subunit). Preliminary analyses suggested that degradation of alpha3 may play a role in promoting release and spread of progeny virions.
authors.affiliation: Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Dallaire
authors.forename: Paola
authors.initials: P
authors.lastname: Blanchette
authors.forename: Peter
authors.initials: P
authors.lastname: Groitl
authors.forename: Thomas
authors.initials: T
authors.lastname: Dobner
authors.forename: Philip E
authors.initials: PE
authors.lastname: Branton
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism*; D017501:Adenovirus E1B Proteins / Q000235:genetics / Q000378:metabolism*; D017504:Adenovirus E4 Proteins / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D044842:Cullin Proteins / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D039422:Integrin alpha3 / Q000378:metabolism*; D008970:Molecular Weight; D011485:Protein Binding; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D040901:Proteomics; D013379:Substrate Specificity; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000378:metabolism
chemical_list: D017501:Adenovirus E1B Proteins; D017504:Adenovirus E4 Proteins; C496871:CUL5 protein, human; D044842:Cullin Proteins; D039422:Integrin alpha3; D021122:Protein Subunits; D044767:Ubiquitin-Protein Ligases