The algD gene encodes NAD-linked GDPmannose dehydrogenase, which is essential for the mucoid phenotype, an important virulence factor expressed by Pseudomonas aeruginosa in cystic fibrosis patients. AlgR, a response regulator controlling mucoidy, is required for high level expression of algD. Inactivation of algR completely abrogates algD expression while mutations immediately downstream of algR affect induction of the algD promoter. In order to examine the nature of genetic elements located downstream of algR, the complete nucleotide sequence of this region was determined. This analysis revealed the presence of two newly identified P. aeruginosa genes with predicted gene products homologous to known porphobilinogen deaminases (HemC) from other organisms, and uroporphyrinogen III cosynthase (HemD) from Escherichia coli. The concerted action of both of these enzymes is essential for the synthesis of heme precursors. Mutations within the region containing the P. aeruginosa homologs of hemC and hemD affect algD promoter activity during growth on nitrate. Furthermore, transcriptional analyses indicated that hemC was cotranscribed with algR at detectable levels in mucoid cells. These results suggest a link between physiological processes dependent on heme and conditions conductive to algD expression and mucoidy.