Phytochelatin synthase genes from arabidopsis and the yeast Schizosaccharomyces pombe

被引:490
作者
Ha, SB
Smith, AP
Howden, R
Dietrich, WM
Bugg, S
O'Connell, MJ
Goldsbrough, PB
Cobbett, CS [1 ]
机构
[1] Univ Melbourne, Dept Genet, Parkville, Vic 3052, Australia
[2] Purdue Univ, Dept Hort & Landscape Architecture, W Lafayette, IN 47907 USA
[3] Peter MacCallum Canc Inst, Trescowthick Res Labs, Melbourne, Vic 3000, Australia
关键词
D O I
10.1105/tpc.11.6.1153
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Phytochelatins (PCs), a family of heavy metal-inducible peptides important in the detoxification of heavy metals, have been identified in plants and some microorganisms, including Schizosaccharomyces pombe, but not in animals. PCs are synthesized enzymatically from glutathione (GSH) by PC synthase in the presence of heavy metal ions. In Arabidopsis, the CAD1 gene, identified by using Cd-sensitive, PC-deficient cad1 mutants, has been proposed to encode PC synthase. Using a positional cloning strategy, we have isolated the CAD1 gene. Database searches identified a homologous gene in S. pombe, and a mutant with a targeted deletion of this gene was also Cd sensitive and PC deficient, Extracts of Escherichia coli cells expressing a CAD1 cDNA or the S. pombe gene catalyzing GSH-dependent, heavy metal-activated synthesis of PCs in vitro demonstrated that both genes encode PC synthase activity. Both enzymes were activated by a range of metal ions. In contrast, reverse transcription-polymerase chain reaction experiments showed that expression of the CAD1 mRNA is not influenced by the presence of Cd, A comparison of the two predicted amino acid sequences revealed a highly conserved N-terminal region, which is presumed to be the catalytic domain, and a variable C-terminal region containing multiple Cys residues, which is proposed to be involved in activation of the enzyme by metal ions. Interestingly, a similar gene was identified in the nematode, Caenorhabditis elegans, suggesting that PCs may also be expressed in some animal species.
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页码:1153 / 1163
页数:11
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