bcl-x sub(L) is the major bcl-x mRNA form expressed during murine development and its product localizes to mitochondria

Most examples of cell death in animals are controlled by a genetic program that is activated within the dying cell. The apoptotic process is further regulated by a set of genes that act as repressors of cell death. Of these, bcl-2 is expressed in a variety of embryonic and postnatal tissues which suggests a critical role for bcl-2 in organogenesis and tissue homeostasis. Surprisingly, mutant mice with targeted disruption of bcl-2 appear normal at birth and complete maturation of lymphoid tissues before succumbing to fulminant lymphopenia and polycystic renal disease by 2-5 weeks of age. This suggests that there may be genes other than bcl-2 that can regulate apoptosis during development. To begin to investigate this possibility, we have cloned and characterized the murine bcl-x gene, whose human counterpart displays striking homology to bcl-2. The predicted murine bcl-x sub(L) gene product exhibits a high level of amino acid identity (97%) to its human counterpart. Just like Bcl-2, the murine bcl-x sub(L) gene product can act as a dominant inhibitor of cell death upon growth factor withdrawal. In addition, the bulk of the bcl-x sub(L) product localizes to the periphery of mitochondria as assessed by a bcl-x sub(L)-tag expression system, suggesting that both Bcl-2 and Bcl-x sub(L) proteins prevent cell death by a similar mechanism. bcl-x sub(L) is the most abundant bcl-x mRNA species expressed in embryonic and adult tissues. The levels of bcl-x sub(L) mRNA appear higher than those of bcl-2 during embryonal development and in several adult organs including bone marrow, brain, kidney and thymus. In addition to bcl-x sub(L), we have identified another form of bcl-x mRNA, bcl-x sub( beta ), that results from an unspliced bcl-x transcript. bcl-x sub( beta ) mRNA is expressed in various embryonic and postnatal tissues. Surprisingly, the expression of bcl-x sub(S) (a negative regulator of programmed cell death) was undetectable by a sensitive S1-nuclease assay and polymerase chain reaction analysis of mouse tissues. Based on its tissue and developmental patterns of expression, it appears that bcl-x may play an important role in the regulation of cell death during development and tissue homeostasis. (DBO)