Copper-dependent control of uptake, translocation and accumulation of cadmium in hyperaccumlator Sedum alfredii.

Cadmium (Cd) is detrimental to plant growth and threatens human health. Here, we investigated the potential for remediation of Cd-contaminated soil with high copper (Cu) background using Cd hyperaccumulator ecotype (HE) Sedum alfredii. We assessed effects of Cu on Cd accumulation, compartmentation and translocation in HE S. alfredii, and compared with those in a related non-accumulator ecotype (NHE). We found that Cu supply significantly induced Cd accumulation in roots and shoots of long-term soil-cultivated HE S. alfredii. A large fraction of root Cd was accumulated in the organelles, but a small fraction was stored in the cell wall. Importantly, Cu addition reduced Cd accumulation in the cell wall and the organelles in root cells. Furthermore, leaf cell capacity to sequestrate Cd in the organelles was greatly improved upon Cu exposure. We also found that genes involving metal transport and cell wall remodeling were distinctly regulated to mediate Cd accumulation in HE S. alfredii. These findings indicate that Cu-dependent decrease of root cell-wall-bound Cd, and stimulation of efflux/influx of organelle Cd transport in root and leaf cells plays a role in the dramatic Cd hyperaccumulation expressed in naturally survived HE S. alfredii.

Arabidopsis COPT1 copper transporter uses a single histidine to regulate transport activity and protein stability.

Copper acquisition and subsequent delivery to target proteins are essential for many biological processes. However, the cellular levels of this trace element must be controlled because of its potential toxicity. The COPT1 protein rich in potential metal-binding amino acids functions in high affinity copper uptake at the plasma membrane of Arabidopsis cells. The functional role of these putative metal-binding residues is largely unknown. Through truncations and site-directed mutagenesis, we identified His43, a single residue within the extracellular N-terminal domain as absolutely critical for copper uptake of COPT1. Substitution of this residue with leucine, methionine or cysteine almost inactivated transport function of COPT1, implying that His43 fails to serves as a copper ligand in the regulation of COPT1 activity. Deletion of all extracellular N-terminal metal-binding residues completely blocked copper-stimulated degradation but did not alter the subcellular distribution and multimerization of COPT1. Although mutation of His43 to alanine and serine retained the transporter activity in yeast cells, the mutant protein was unstable and degraded in the proteasome in Arabidopsis cells. Our results demonstrate a pivotal role for the extracellular residue His43 in high affinity copper transport activity, and suggest common molecular mechanisms for regulating both metal transport and protein stability of COPT1.