Isolation and characterization of a cDNA that encodes maize uroporphyrinogen III methyltransferase, an enzyme involved in the synthesis of siroheme, which is prosthetic group of nitrite reductase.

Abstract

A full-length cDNA clone, pZmSUMT1, encoding an S-adenosyl-L-methionine-dependent uroporphyrinogen III C-methyltransferase (SUMT; EC 2.1.1.107) of maize was isolated from a root cDNA library. pZmSUMT1 had an insert of 1.7 kb and the amino acid sequence deduced from the open reading frame of the cDNA was similar to that of SUMT from various bacteria and also to the SUMT catalytic region of siroheme synthase (cysG) from Escherichia coli. Overproduction of ZmSUMT1 in a cysG mutant of E. coli eliminated the requirement of the strain for cysteine. Transcripts for ZmSUMT1 accumulated rapidly in both rots and leaves in response to the addition of nitrate to the culture medium. The effects of biochemical inhibitors on the nitrate-dependent induction of the gene for ZmSUMT1 coincided with the effects on the genes for other nitrate-assimilatory enzymes, nitrate reductase and nitrite reductase. An import experiment in vitro suggested that the gene product might be located in plastids. The results together indicate that ZmSUMT1 might be involved in the synthesis of siroheme, a prosthetic group of nitrite reductase, and that the expression of its gene is co-regulated with that of other nitrate-assimilatory genes.