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Nucleic Acids Research, 1989, Vol. 17, No. 24 10231-10241
© 1989

Articles

Increased Sp1 binding mediates erythroid-specific overexpression of a mutated (HPFH) {gamma}-globulin promoter

Antonella Ronchi*, Silvia Nicolis, Claudio Santoro1 and Sergio Ottolenghi

Dipartimento di Genetica e di Biologia dei Microrganismi, Universita di Milano Italy 1Dipartimento di Genetica, Biologia e Chimica Medica, Università di Torino Italy

*To Whom Correspondance should be addressed

Received October 2, 1989. Revised November 22, 1989. Accepted November 22, 1989.

The ~198 T->C mutation in the promoter of the A{gamma}-globin gene increases 20–30 fold the expression of this gene in adult erythroid cells of patients (Hereditary Persistence of Fetal Hemoglobin, HPFH). We show here that this mutation creates a strong binding site, resembling a CACCC box, for two ubiquitous nuclear proteins, one of which is Spl. The mutated promoter is four to five-fold more efficient than a normal {gamma}globin promoter in driving expression of a CAT reporter plasmid when transfected into erythroid cells. The overexpression of the mutant is abolished by the introduction of an additional mutation disrupting the new binding site. No overexpression of the mutant is observed in non-erythroid cells, indicating that the ubiquitous factors bound on the mutated sequence must cooperate with erythroid specific factors.


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