In a variety of physiological settings, cells are eliminated by apoptosis-a genetically encoded process of cellular suicide. Bak, a member of the Bcl-2 protein family, accelerates apoptosis by an unknown mechanism. We have found a novel cDNA encoding a 101-amino acid protein that possesses a Bak-like sequence in our full-length cDNA bank and termed it Bak-like. This protein shares the conserved domains BH1 and BH2 with other pro-apoptotic proteins, but lacks the BH3 domain. Database searches identified this gene on chromosome 6, which could account for the cloned bak and bak-like transcripts by alternative splicing. Bak-like is expressed in a wide variety of tissues. Bak-like is different from bak by Southern blots using probes with or without homology to bak. Despite the loss of the BH3 sequence, bak-like did enhance apoptosis, but was less potent than bak. Confocal microscopy of HeLa cells revealed that EGFP-Bak-like was located diffusely throughout the cytosol. However, upon induction of apoptosis, EGFP-Bak-like redistributed into a punctuate pattern, colocalizing with mitochondria. Like bak, the bak-like gene product directly enhanced apoptotic cell death following an appropriate stimulus.