Characterization of a distal 5'-flanking region (-2010/-630) of human GM-CSF

The 5'-flanking region of the human GM-CSF gene was subcloned from the phagic clone lambda J1-16 to detect cis-elements involved in GM-CSF constitutive expression. We determined and sequenced an uncharacterized promoter region of 1381 bp (-2010 to -630), named pPF2000. Putative binding sites of several transcriptional factors were found. Progressive deletion mutants of the PF2000 were analyzed by measuring the linked CAT activities, in constitutive (5637) and inducible (PEU) GM-CSF-producing cells. A positive distal sequence (268 bp), between -2010 and -1742, responsible for the high constitutive expression of GM-CSF in 5637 carcinoma cell line was found.