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Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles
Published on: December 31, 2019
Genetic characterization of oligopeptide uptake systems in Sinorhizobium meliloti
Joaquina Nogales1, Socorro Muñoz, José Olivares
1Dpto. Microbiología del Suelo y Sistemas Simbióticos, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Granada, Spain.
Abstract:
The genetic characterization of three ABC transport systems involved in oligopeptide uptake by Sinorhizobium meliloti is reported. Oligopeptide permease (Opp) encoded by the pSymB oppABCD operon, is required for uptake of tetrapeptides and certain tripeptides like 3Ala and bialaphos. The chromosomally encoded dipeptide permease (Dpp1), also able to import the toxic tripeptide bialaphos, is required for utilization of dipeptides and tripeptides like 3Gly and GlyGlyAla, with minor importance for utilization of 3Ala and tetrapeptides. The ttp (tri and tetrapeptide uptake) operon, encodes a third ABC system (Ttp) unable of transporting bialaphos and with minor role in the utilization of tetrapeptides and tripeptides like 3-Ala. Despite the overlapping substrate specificities of these ABC transporters, the corresponding gene operons displayed distinct expression profiles: dpp1 showed high constitutive expression levels under all conditions tested, in contrast to the low expression levels of ttp, whereas opp was maximally expressed upon entry into stationary phase. Nevertheless, complex interactions among the three systems at the transcriptional level were observed: opp was negatively autoregulated via OppA and positively regulated via DppA1, whereas dpp1 seems negatively autoregulated via DppA1. The expression of both opp and dppl was reduced in Ttp mutants. The ABC transport systems characterized in this work are not essential for the establishment of nitrogen-fixing symbiosis with alfalfa.
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