CopM is a novel copper-binding protein involved in copper resistance in Synechocystis sp. PCC 6803

Abstract

Copper resistance system in the cyanobacterium Synechocystis sp. PCC 6803 comprises two operons, copMRS and copBAC, which are expressed in response to copper in the media. copBAC codes for a heavy-metal efflux-resistance nodulation and division (HME-RND) system, while copMRS codes for a protein of unknown function, CopM, and a two-component system CopRS, which controls the expression of these two operons. Here, we report that CopM is a periplasmic protein able to bind Cu(I) with high affinity (KD ~3 × 10-16). Mutants lacking copM showed a sensitive copper phenotype similar to mutants affected in copB, but lower than mutants of the two-component system CopRS, suggesting that CopBAC and CopM constitute two independent resistance mechanisms. Moreover, constitutive expression of copM is able to partially suppress the copper sensitivity of the copR mutant strain, pointing out that CopM per se is able to confer copper resistance. Furthermore, constitutive expression of copM was able to reduce total cellular copper content of the copR mutant to the levels determined in the wild-type (WT) strain. Finally, CopM was localized not only in the periplasm but also in the extracellular space, suggesting that CopM can also prevent copper accumulation probably by direct copper binding outside the cell. We have characterized a new periplasmic copper-binding protein (CopM) that is part of the copper resistance mechanism in Synechocystis sp. PCC 6803. We have showed that CopM is able to bind both Cu(I) and Cu(II) and to confer resistance to copper. Furthermore, CopM is found not only in the periplasm but also in the extracellular space and is able to reduce total copper contents in Synechocystis sp. PCC 6803.