The t (12; 21) translocation is present in up to 30% of childhood B-cell acute lymphoblastic leukemias and fuses a potential dimerization motif from the ets-related factor TEL to the N terminus of AML1. The t (12; 21) translocation encodes a 93-kDa fusion protein that localizes to a high-salt-and detergent-resistant nuclear compartment. This protein binds the enhancer core motif, TGTGGT, and interacts with the AML-1-binding protein, core-binding factor beta. Although TEL/AML-1B retains the C-terminal domain of AML-1B that is required for transactivation of the T-cell receptor beta enhancer, it fails to activate transcription but rather inhibits the basal activity of this enhancer. TEL/AML-1B efficiently interferes with AML-1B-dependent trans-activation of the T-cell receptor beta enhancer, and coexpression of wild-type TEL does not reverse this inhibition. The N-terminal TEL helix-loop-helix domain is essential for TEL/AML-1B-mediated repression. Thus, the t (12; 21) fusion protein dominantly interferes with AML-1B-dependent transcription, suggesting that the inhibition of expression of AML-1 target genes is critical for B-cell leukemogenesis.