Epstein-barr virus transcription factor binding assay

Abstract

The invention provides methods and compositions for identifying pharmacological agents useful in the diagnosis or treatment of disease associated with the expression of a gene modulated by a transcription complex containing at least CBF1 and a transcription factor. The methods involve combining CBF1, the Epstein-Barr virus transcription factor EBNA2 or cellular homolog thereof, and a candidate pharmacological agent. This mixture is incubated under conditions whereby, but for the presence of the candidate pharmacological agent, CBF1 binds EBNA2. The absence of selective binding indicates that the candidate pharmacological agent is capable of selectively modulating the expression of a gene dependent on CBF1-transcription factor binding. The mixture may further comprise a CBF1 binding element including the nucleotide sequence: C-G-T-G-G-G-A-A. This mixture is incubated under conditions whereby, but for the presence of the candidate pharmacological agent, a transcription complex comprising both polypeptides bound, directly or indirectly to the nucleic acid is formed. The methods are particularly suited to high-throughput screening where one or more steps are performed by a computer controlled electromechanical robot comprising an axial rotatable arm and said solid substrate is a portion of a well of a microtiter plate.