期刊
METALLOMICS
卷 5, 期 12, 页码 1634-1643出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c3mt00204g
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资金
- Programa de Apoyo a Proyectos de Investigacion e Inovacion Tecnologica (PAPIIT Grant) [IN201112]
- Consejo Nacional de Ciencia y Tecnologia (Mexico) [CVU 269108, 179133]
The ubiquitous Cation Diffusion Facilitator proteins (CDF) play a key role in maintaining the cellular homeostasis of essential metal ions. Previous neighbor-joining phylogenetic analysis classified CDF proteins into three substrate-defined groups: Zn2+, Fe2+/Zn2+ and Mn2+. These studies were unable to discern substrate-defined clades for Ni2+, Co2+, Cd2+ and Cu2+ transporters, despite their existence in this family. In this study we improved the accuracy of this previous functional classification using a phylogenomic approach based on a thorough maximum-likelihood phylogeny and the inclusion of recently characterized CDF transporters. The inference of CDF protein function predicted novel clades for Zn2+, Fe2+, Cd2+ and Mn2+. The Ni2+/Co2+ and Co2+ substrate specificities of two clades containing uncharacterized proteins were defined through the functional characterization of nepA and cepA metal inducible genes which independently conferred Ni2+ and Co2+ resistances to Rhizobium etli CFN42 and increased, respectively, N-i2+/Co2+ and Co2+ resistances to Escherichia coli. Neither NepA nor CepA confer Zn2+, Fe2+ and Mn2+ resistances. The ability of NepA to confer Ni2+/Co2+ resistance is dependent on clade-specific residues Asn(88) and Arg(197) whose mutations produce a non-functional protein.
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