The distal locus control region (LCR) is required for high-level expression of 
the complex of genes (HBBC) encoding the beta-like globins of mammals in 
erythroid cells. Several major DNase hypersensitive sites (HSs 1-5) mark the 
LCR. Sequence conservation and direct experimental evidence have implicated 
sequences within and between the HS cores in function of the LCR. In this report 
we confirm the mapping of a minor HS between HS3 and HS4, called HS3.2, and show 
that sequences including it increase the number of random integration sites at 
which a drug resistance gene is expressed. We also show that nuclear proteins 
including GATA1 and Oct1 bind specifically to sequences within HS3.2. However, 
the protein Pbx1, whose binding site is the best match to one highly conserved 
sequence, does not bind strongly. GATA1 and Oct1 also bind in the HS cores of 
the LCR and to promoters in HBBC. Their binding to this minor HS suggests that 
they may be used in assembly of a large complex containing multiple regulatory 
sequences.