Copper-finger protein of Sp1: the molecular basis of copper sensing

The cellular copper level is strictly regulated since excessive copper is harmful to cells. It has been proposed that the expression of copper transport protein hCtr1 is transcriptionally regulated by specificity protein 1 (Sp1) in response to the cellular copper level. However, it is not known how Sp1, a zinc-finger-protein (ZFP), can sense copper ions in cells. Here we found that Sp1 demonstrates high binding affinity to cuprous ions, even stronger than Cu-Atox1 binding. Cu(i) can displace Zn(ii) in Sp1, resulting in a well-folded 'Copper-Finger-Protein' (CFP). Although only very little structural alteration occurs upon copper binding, CFP cannot recognize the promoter of hCtr1, therefore copper binding interrupts the transcription. This result indicates that, in addition to apo-to-holo alteration, metal substitution can also lead to transcriptional switch in metal sensing. This work provides insight into the copper sensing mechanism of Sp1 at the molecular level.