title: Characterization of bovine adenovirus type 3 E1 proteins and isolation of E1-expressing cell lines.
abstract: Like human adenovirus type 5 (HAV5), bovine adenovirus type 3 (BAV3) early region 1 (E1) consists of E1A and E1B transcriptional units. In order to characterize BAV3 E1 proteins and to isolate a cell line of bovine origin that expresses BAV3 E1, polyclonal antibodies specific to E1A, E1B-157R, and E1B-420R were raised in rabbits. BAV3 E1A, E1B-157R, and E1B-420R were identified as 40, 17, and 47 kDa proteins, and had a half-life of 45-60 min, and 4-6 and 4-6 h, respectively. It appeared that E1A and E1B-157R were phosphorylated at the serine/threonine residues, whereas, E1B 420R was phosphorylated at both the serine/threonine and tyrosine residues. Three cell lines, MDBK-221 (Madin Darby bovine kidney (MDBK) transfected with BAV3 E1), FBK-34 (primary fetal bovine kidney (FBK) cells transfected BAV3 E1), and FBRT-HE1 (bovine fetal retinal (FBRT) cells transfected with HAV5 E1) were isolated and characterized for E1 expression. FBK-34 or FBRT-HE1 supported the replication of an E1A-deleted BAV3 (BAV3DeltaE1AE3) to approximately 1-2 x 10(8) PFU/ml, whereas, the virus titers in MDBK-221 were approximately 10(7) PFU/ml. These cell lines will be useful in generating and growing BAV3 E1-deleted recombinants, and also for studying E1 protein interactions with a number of cellular and/or viral proteins.
authors.affiliation: Department of Veterinary Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, Indiana, 47907, USA.
authors.forename: Alberto L
authors.initials: AL
authors.lastname: van Olphen
authors.forename: Suresh K
authors.initials: SK
authors.lastname: Tikoo
authors.forename: Suresh K
authors.initials: SK
authors.lastname: Mittal
mesh_terms: D017498:Adenovirus E1 Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017500:Adenovirus E1A Proteins / Q000737:chemistry / Q000378:metabolism; D017501:Adenovirus E1B Proteins / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D006207:Half-Life; D017921:Mastadenovirus / Q000378:metabolism*; D008970:Molecular Weight; D010766:Phosphorylation; D012694:Serine / Q000737:chemistry; D014162:Transfection
chemical_list: D017498:Adenovirus E1 Proteins; D017500:Adenovirus E1A Proteins; D017501:Adenovirus E1B Proteins; D012694:Serine