Date of this Version
The Journal of Biological Chemistry, VOL. 284, NO. 7, pp. 4112–4122, February 13, 2009
Copper is an essential yet toxic trace element. The Ctr1 family of proteins plays a critical role for copper uptake in eukaryotes. However, the mechanisms of action of Ctr1 are largely unknown. Our previous data demonstrated that copper transport induces conformational changes in the cytosolic C terminus of the yeast Saccharomyces cerevisiae Ctr1. To define the physiological significance of this molecular event and gain better insights into the mechanism of Ctr1-mediated copper uptake, we have characterized the functional roles of the Ctr1 C terminus.ACtr1 mutant lacking the entire C-terminal cytosolic tail is functional in high affinity copper uptake; however, yeast cells expressing this mutant are extremely sensitive to excess copper. Toxic copper uptake is not attributed to elevated expression or distinct subcellular localization of this mutant as compared with wild type Ctr1. Further characterization of the function of Ctr1 containing deletions or site-directed mutations at theCterminus indicates a structural role for the Cterminus in controlling Ctr1 activities. In response to excess copper, Ctr1- mediated copper transport is rapidly blocked in a C terminusdependent mechanism associated with direct binding of copper. We propose that conformational changes in the cytosolic tail of yeast Ctr1 by copper sensing within this domain lead to the inhibition of Ctr1-mediated copper transport. These data suggest a new regulatory mechanism by which yeast cells maintain homeostatic copper acquisition.