The cyanobacterium Synechococcus sp. PCC 7942 contains two endogenous, genetically cryptic plasmids pANS and pANL, respectively. Characterization of the 3.8 kb Ba6 BamHI fragment of pANL identified three open reading frames which were transcriptionally regulated by sulfur availability and the protein CysR. One of these genes, designated srpG, encodes a protein which exhibits 67% amino acid identity to the Escherichia coli enzyme O-acetyl-l-serine (thiol)-lyase A. Overlapping the 3 end of srpG is a second gene, designated srpH, which encodes a protein with similarity to the amino-terminal region of serine acetyltransferase enzymes. DNA hybridization results indicate that there is a second copy of srpG in Synechococcus sp. PCC 7942, which is consistent with previous isoenzyme studies on O-acetyl-l-serine (thiol)-lyase in cyanobacteria. The introduction of srpG and srpH into E. coli cysKcysM and cysE mutant strains, respectively, results in the complementation of the lesion in cysteine biosynthesis. Additionally, the E. coli cysK cysM strain containing srpG is able to utilize sulfate more efficiently than thiosulfate, indicating that SrpG is probably a type A O-acetyl-l-serine (thiol)-lyase. The possible function of these genes in the adaptation of cyanobacteria to sulfur stress is discussed.