This review summarizes the indispensable requirement of transcription factor PU.1 during hematopoietic cell fate decisions and how the function of PU.1 can be modulated by protein–protein interactions with additional factors. The mutual negative regulation between PU.1 and GATA-1 is detailed within the context of normal and leukemogenic hematopoiesis and the concept of 'differentiation therapy' to restore normal cellular differentiation of leukemic cells is discussed.