We have determined the chromosomal location of the human genes coding for the alpha 3 and beta 3 subunits of soluble guanylyl cyclase (GC-S). The study was performed by in situ hybridization of human metaphase spreads with two human cDNA probes, containing the coding sequences of the GC-S alpha 3 and beta 3 subunits, respectively. Each probe gave a strong specific signal on chromosome 4 at the 4q31.3-4q33 region, with the maximal signal in the 4q32 band. The colocalization of both genes in 4q32 represents an interesting feature for the coordinated regulation of expression of both GC-S subunits.