title: Expression of the adenovirus E1B 175R protein and its association with membranes of Escherichia coli.
abstract: The E1B 175-amino-acid (175R) protein of adenovirus 2 is required for cellular transformation of primary cells and establishing cell morphology in lytically infected cells. To investigate the biochemical function of this protein, we constructed a bacterial expression vector (pKHB1-T) to produce the 175R protein in sufficient amounts for purification and biochemical analysis. On the basis of DNA sequencing, gel electrophoresis, and immunoblot analysis, the pKHB1-T-encoded 175R protein appears to be identical to that expressed transiently in mammalian or adenovirus-transformed cells. The bacterially produced viral protein was also found to be quite stable and without any modifications. Partial purification of the pKHB1-T-encoded protein revealed that the majority of its associates with the inner membrane of the bacterial cell. This, together with the possibility of the 175R protein containing an N-terminal amphipathic alpha-helix as a potential translocation signal, suggests that there may be a common mechanism of protein transport operating in both eucaryotic and procaryotic systems.
authors.affiliation: Department of Molecular Biology, Cleveland Clinic Foundation, Ohio 44195.
authors.forename: B
authors.initials: B
authors.lastname: Dalie
authors.forename: J L
authors.initials: JL
authors.lastname: Ko
authors.forename: D M
authors.initials: DM
authors.lastname: Wang
authors.forename: K
authors.initials: K
authors.lastname: Babich
authors.forename: A C
authors.initials: AC
authors.lastname: Banerjee
authors.forename: M L
authors.initials: ML
authors.lastname: Harter
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D017501:Adenovirus E1B Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004926:Escherichia coli / Q000737:chemistry*; D005347:Fibroblasts; D015870:Gene Expression; D005822:Genetic Vectors; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
chemical_list: D017501:Adenovirus E1B Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins