Bak can accelerate chemotherapy-induced cell death independently of its heterodimerization with Bcl-X_L and Bcl-2

Abstract

Bak has been shown to both promote apoptosis and to inhibit cell death while two other members of the Bcl-2 family of proteins, Bcl-X_L and Bcl-2 delay apoptosis induced by various stimuli including chemotherapeutic agents. We generated clones with stable expression of Bak wild-type (wt) and Bak with its BH3 (Δ78-86) domain deleted (ΔBH3) in FL5.12 cells or FL5.12 cells expressing either Bcl-X_L or Bcl-2 to determine if Bak could accelerate apoptosis and antagonize the death repressor activity of Bcl-X_L and Bcl-2 during chemotherapy-induced apoptosis. We found that Bak accelerated cell death in FL5.12 cells treated with etoposide, fluorouracil or taxol. In FL5.12 cells expressing Bcl-X_L and Bak wt or Bak ΔBH3, both Bak wt or Bak ΔBH3 were able to antagonize the protective effect of Bcl-X_L when treated with etoposide or fluorouracil. Bak wt or Bak ΔBH3 were also able to abrogate the protective effect of Bcl-2 in cells expressing Bcl-2 and Bak wt or Bak ΔBH3 when challenged by etoposide or fluorouracil. Immunoprecipitation studies revealed that deletion of BH3 disrupted heterodimerization between Bak and Bcl-X_L and that both Bak wt and Bak ΔBH3 failed to interact with Bcl-2. These results demonstrate that Bak does not require its BH3 domain to promote apoptosis in stably transfected cells. Furthermore, Bak can accelerate chemotherapy-induced cell death independently of its heterodimerization with Bcl-X_L and Bcl-2.