Characterization of adenovirus type 5 insertion and deletion mutants encoding altered DNA binding proteins.
We have introduced insertion and deletion mutations in the cloned DNA binding protein (DBP) gene of adenovirus type 5. The mutated DBP genes were subsequently introduced in the viral genome by a combination of in vitro and in vivo methods. The resulting mutant viruses were tested for their viability in human 293 cells and an initial characterization of these viruses was performed. Viable mutants with insertions in the carboxyl-terminal portion of the gene could not be obtained. In contrast, a number of viable mutants were constructed that contained insertions or deletions in the amino-terminal half of DBP. Several of these, which covered the region between amino acid (aa) residues 39 and 81, were phenotypically wild type, implying that this segment is completely dispensable for DBP function. However, mutations altering the region encompassed by aa 2-38 were, at least, partially defective suggesting that this region is important for full activity of the protein.
Vos
H L
HL
Laboratory for Physiological Chemistry, State University of Utrecht, the Netherlands.
Brough
D E
DE
Van der Lee
F M
FM
Hoeben
R C
RC
Verheijden
G F
GF
Dooijes
D
D
Klessig
D F
DF
Sussenbach
J S
JS
D000260:Adenoviruses, Human / Q000235:genetics* / Q000254:growth & development; D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping; D014162:Transfection
D004279:DNA, Viral; D004268:DNA-Binding Proteins