title: Functional characterization of the CDF transporter SMc02724 (SmYiiP) in Sinorhizobium meliloti: Roles in manganese homeostasis and nodulation.
abstract: In bacteria, membrane transporters of the cation diffusion facilitator (CDF) family participate in Zn(2+), Fe(2+), Mn(2+), Co(2+) and Ni(2+) homeostasis. The functional role during infection processes for several members has been shown to be linked to the specificity of transport. Sinorhizobium meliloti has two homologous CDF genes with unknown transport specificity. Here we evaluate the role played by the CDF SMc02724 (SmYiiP). The deletion mutant strain of SmYiiP (ΔsmyiiP) showed reduced in vitro growth fitness only in the presence of Mn(2+). Incubation of ΔsmyiiP and WT cells with sub-lethal Mn(2+) concentrations resulted in a 2-fold increase of the metal only in the mutant strain. Normal levels of resistance to Mn(2+) were attained by complementation with the gene SMc02724 under regulation of its endogenous promoter. In vitro, liposomes with incorporated heterologously expressed pure protein accumulated several transition metals. However, only the transport rate of Mn(2+) was increased by imposing a transmembrane H(+) gradient. Nodulation assays in alfalfa plants showed that the strain ΔsmyiiP induced a lower number of nodules than in plants infected with the WT strain. Our results indicate that Mn(2+) homeostasis in S. meliloti is required for full infection capacity, or nodule function, and that the specificity of transport in vivo of SmYiiP is narrowed down to Mn(2+) by a mechanism involving the proton motive force.
authors.affiliation: Instituto de Investigación Médica Mercedes y Martín Ferreyra (INIMEC), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Córdoba, Córdoba, Argentina. Electronic address: draimunda@immf.uncor.edu.
authors.forename: Daniel
authors.initials: D
authors.lastname: Raimunda
authors.affiliation: Instituto de Investigación Médica Mercedes y Martín Ferreyra (INIMEC), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Córdoba, Córdoba, Argentina.
authors.forename: Graciela
authors.initials: G
authors.lastname: Elso-Berberián
keywords: Cation diffusion facilitator; Manganese efflux; Manganese homeostasis; Sinorhizobium meliloti