Coactivator CBP (CREB-binding protein) has been implicated in the regulation of transcription for all signal transducer and activator of transcription factors (STATs); however, the mechanism remains unclear. Using yeast two-hybrid screening and immunoprecipitation techniques, we investigated the direct interaction of CBP with STAT4 and STAT6. The full-length CBP and five fragments of CBP (residues 1-436, 529-1200, 1-697, 967-1574 and 1678-2175) were constructed using pGBKT7 vectors, while STAT4, STAT6 and N-terminal deleted STAT4 were constructed using pGADT7 vectors. It was found that STAT4, but not STAT6, interacted directly with the 1678-2175 fragment of CBP containing the ZZ, TAZ2 and SID domain. The N-terminal of STAT4 plays an important role in this interaction since N-terminal deleted STAT4 failed to bind to any CBP fragment. The results were confirmed by immunoprecipitation using HA-tagged STAT4 or STAT6 and c-Myc tagged CBP. This work will contribute to our understanding of the mechanisms of Th cytokine imbalance.