CHARACTERIZATION OF A SUBSET OF THE BASIC-HELIX-LOOP-HELIX-PAS SUPERFAMILY THAT INTERACTS WITH COMPONENTS OF THE DIOXIN SIGNALING PATHWAY

In an effort to better understand the mechanism of toxicity of 2,3,7,8 -tetrachlorodibenzo-p-dioxin, we employed an iterative search of human expressed sequence tags to identify novel basic-helix-loop-helix-PAS (bHLH-PAS) proteins that interact with either the Ah receptor (AHR) or the Ah receptor nuclear translocator (ARNT). We characterized five ne w ''members of the PAS superfamily,'' or MOPs 1-5, that are similar in size and structural organization to the AHR and ARNT. MOPs 1-4 have N -terminal bHLH and PAS domains and C-terminal variable regions. MOP5 c ontained the characteristic PAS domain and a variable C terminus; it i s possible that the cDNA contains a bHLH domain, but the entire open r eading frame has yet to be completed. Coimmunoprecipitation studies, y east two-hybrid analysis, and transient transfection experiments demon strated that MOP1 and MOP2 dimerize with ARNT and that these complexes are transcriptionally active at defined DNA enhancer sequences in viv o. MOP3 was found to associate with the AHR in vitro but not in vivo. This observation, coupled with the fact that MOP3 formed tighter assoc iations with the 90-kDa heat shock protein than the human AHR, suggest s that MOP3 may be a conditionally active bHLH-PAS protein that requir es activation by an unknown ligand. The expression profiles of the AHR , MOP1, and MOP2 mRNAs, coupled with the observation that they all sha re ARNT as a common dimeric partner, suggests that the cellular pathwa ys mediated by MOP1 and MOP2 may influence or respond to the dioxin si gnaling pathway.