We have analyzed the genomic interval between the MHC-linked heat shock protein 70 (Hsp70) genes Hsp70-2 and Hsp70-3 in the rat. The distance between the transcription start sites of both genes which are organized in a head-to-head orientation is 604 bp. Unlike Hsp70-2 the 5' untranslated region of Hsp70-3 is interrupted by an intron of about 3.9 kb. The Hsp70-3 promoter lacks a TATA box and heat shock elements, but contains two CRE elements, one Sp1-GC and two PuF binding sites. Analysis of the Hsp70-2 promoter by CAT assays reveals positive and negative regulatory elements extending into the Hsp70-3 gene. (C) 1998 Elsevier Science B.V.