Elf-1 and Stat5 bind to a critical element in a new enhancer of the human interleukin-2 receptor α gene

The interleukin 2 receptor α-chain (IL-2Rα) gene is a key regulator of lymphocyte proliferation. IL-2Rα is rapidly and potently induced in T cells in response to mitogenic stimuli. Interleukin 2 (IL-2) stimulates IL-2Rα transcription, thereby amplifying expression of its own high-affinity receptor. IL-2Rα transcription is at least in part controlled by two positive regulatory regions, PRRI and PRRII. PRRI is an inducible proximal enhancer, located between nucleotides -276 and -244, which contains NF-κB and SRE/CArG motifs. PRRII is a T-cell-specific enhancer, located between nucleotides -137 and -64, which binds the T-cell-specific Ets protein Elf-1 and HMG-1(Y) proteins. However, none of these proximal regions account for the induction of IL-2Rα transcription by IL-2. To find new regulatory regions of the IL-2Rα gene, 8.5 kb of the 5' end noncoding sequence of the IL-2Rα gene have been sequenced. We identified an 86-nucleotide fragment that is 90% identical to the recently characterized murine IL-2-responsive element (mIL-2rE). This putative human IL-2rE, designated PRRIII, confers IL- 2 responsiveness on a heterologous promoter. PRRIII contains a Stat protein binding site that overlaps with an EBS motif (GASd/EBSd). These are essential for IL-2 inducibility of PRRIII/CAT reporter constructs. IL-2 induced the binding of Stat5a and b proteins to the human GASd element. To confirm the physiological relevance of these findings, we carried out in vivo footprinting experiments which showed that stimulation of IL-2Rα expression correlated with occupancy of the GASd element. Our data demonstrate a major role of the GASd/EBSd element in IL-2Rα regulation and suggest that the T- cell-specific Elf-1 factor can serve as a transcriptional repressor.