Isolation of the gene HEM4 encoding uroporphyrinogen III synthase in Saccharomyces cerevisiae

Abstract

We have isolated a genomic DNA fragment that complements the yeast temperature-sensitive cyt mutation, causing respiratory deficiency and accumulation of porphyrins (Sugimura et al., 1966). Partial DNA sequencing of the complementing region and search for similarity in the DNA and protein databases revealed that (1) the gene had been previously isolated by complementation of the mutation ts2326 (Langgut et al., 1986; accession number X04694), and (2) it encodes a protein with 18-23% identity to uroporphyrinogen III synthases from different sources. This enzyme catalyses the fourth step in the heme biosynthetic pathway and we named its gene HEM4. A hem4 delta disruption mutation was constructed which had phenotypes identical to the cyt mutation. Biochemical analysis confirmed the absence of uroporphyrinogen III synthase activity in both hem4 delta and cyt mutant strains.