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The relationship between region E1a and E1b of human adenoviruses in cell transformation
Authors:P van den Elsen  S de Pater  A Houweling  J van der Veer  A van der Eb
Affiliation:Department of Medical Biochemistry, Sylvius Laboratories, Wassenaarseweg 72, 2333 AL LeidenThe Netherlands
Abstract:Baby rat kidney (BRK) cells were transfected either with intact region E1 DNA of adenovirus type 5 (Ad5) or with mixtures of DNA fragments containing the separated E1a and E1b regions. The results showed that mixtures of regions E1a and E1b transform with a similar efficiency as intact region E1. DNA fragments containing region E1b alone have no detectable transforming activity in primary BRK cells nor in established rat cell lines. When region E1a and Ad5 was combined with region E1b and Ad12 complete transformation was also obtained. Characterization of the cell lines transformed by separated E1a and E1b regions have led to the following conclusions: (1) Expression of region E1b is not dependent on specific linkage to region E1a as it occurs in the intact E1 region. (2) Region E1b is normally expressed into the corresponding major adenovirus T antigens (65,000 and 19,000 Mr with region E1b of Ad5; 60,000 and 19,000 Mr with E1b or AD12). (3) Region E1b of Ad12 can be activated by region E1a of Ad5 indicating that the Ela regions of both serotypes are functionally similar in transformation. (4) Cell lines containing region E1b of Ad5 are weakly oncogenic in nude mice whereas cells containing E1b of Ad12 are highly oncogenic in nude mice, indicating that the degree of oncogenicity is determined by region E1b.
Keywords:Transformation by adenovirus 5 and 12  DNA transfection  oncogenicity in nude mice  Ad  adenovirus  bp  base pairs  BRK  baby rat kidney  DMSO  dimethyl sulphoxide  HeBS  HEPES-buffered saline  p  plasmid  PBS  phosphate-buffered saline
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