Mouse mastocytoma P815 expresses several distinct tumor rejection antigens recognized by syngeneic cytolytic T lymphocytes (CTL). Two of these tumor rejection antigens, P815A and P815B, are encoded by gene P1A, the sequence of which was reported previously. Tumor cell variants having lost one or both of these antigens were isolated by in vitro selection with CTL and also by collecting tumor cells that progressed in vivo after escaping a nearly complete immune rejection process. The structure of gene P1A in these antigen‐loss variants was examined. Several A⁻B⁻ variants presented a complete or partial deletion of the gene. One variant that had lost only antigen A displayed a point mutation in the first exon. Peptides were synthesized that corresponded to the normal sequence located in the region of this point mutation. They sensitized target cells to both anti‐A and anti‐B CTL. The homologous peptide encoded by the mutated gene of the P815 A⁻B⁺ variant sensitized cells only to anti‐B CTL. We conclude that anti‐A and anti‐B CTL recognize on the same peptide two distinct epitopes that are affected differently by the mutation.