Construction and functional characterization of mutants of the bovine leukaemia virus trans-activator protein p34tax.
The p34tax protein [p38tax, p34, p38(XBL), XBL-I] of bovine leukaemia virus (BLV) activates transcription from the BLV long terminal repeat (LTR) promoter. To analyse the functional properties of this protein, inframe insertions and internal deletions were systematically introduced in a plasmid-encoded copy of the p34tax gene. The abilities of wild-type and mutant genes to activate gene expression from the LTR promoter linked to the chloramphenicol acetyltransferase gene and to inhibit trans-activation by the wild-type protein were studied. The trans-activating activity of 14 of the 18 mutants tested was completely abolished, but four mutants each containing a lesion in the internal portion of the polypeptide retained activity. Taken together, these results suggest the presence of an internal region of the polypeptide where structural integrity is less strictly required for the functional activity of this protein. Among the mutants incompetent in the transactivation assay, only two with mutations in the N-terminal region of the polypeptide inhibited transactivation by the wild-type protein in a dose-dependent manner. These results facilitate understanding of the physiological function of the tax protein family.
Sakurai
M
M
Biophysics Laboratory, National Institute of Animal Health, Ibaraki, Japan.
Taneda
A
A
Nagoya
H
H
Sekikawa
K
K
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D016356:Gene Products, tax / Q000235:genetics*; D001909:Leukemia Virus, Bovine / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D010957:Plasmids; D015183:Restriction Mapping; D012756:Sheep
D016356:Gene Products, tax