HTLV-I Tax self-association in optimal trans-activation function.
HTLV-I Tax protein is a potent transcriptional activator of viral and cellular genes. Tax does not bind DNA directly but interacts through protein-protein contact with host cell factors that recognize the viral long terminal repeat (LTR). Domains within Tax needed for protein-protein interaction have not been fully characterized. In studying transcriptional function in yeast cells, we unexpectedly found that Tax functions optimally not as a monomer, but as a homodimer. Here we have used the one hybrid and two hybrid genetic approaches in yeast to investigate the region(s) within Tax necessary for self-association. Dimer formation was also confirmed biochemically by using electrophoretic mobility shift (EMSA) and supershift assays. Twenty two Tax point mutants were utilized to map relevant residues. Genetic results from this series of mutants revealed that a necessary region for dimerization is contained within a previously characterized zinc finger domain. Two loss-of-function Tax mutants, each poorly active when assayed individually, were found to have complementing activity when co-expressed together. This genetic complementation suggests a mechanism fortrans-activation resulting from simultaneous but non-identical contact with a responsive target by each of two Tax monomers in a dimer.
Jin
D Y
DY
Molecular Virology Section, Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-0460, USA.
Jeang
K T
KT
D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression / Q000235:genetics; D016356:Gene Products, tax / Q000737:chemistry / Q000378:metabolism*; D005816:Genetic Complementation Test; D008956:Models, Chemical; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D017354:Point Mutation / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics; D015533:Transcriptional Activation / Q000235:genetics*; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D016356:Gene Products, tax; D014764:Viral Proteins; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase