Characterization of outer membrane iron uptake pathways in the model cyanobacterium Synechocystis sp. PCC 6803.

Cyanobacteria are foundational drivers of global nutrient cycling with high intracellular iron (Fe) requirements. Fe is found at extremely low concentrations in aquatic systems however, and the ways in which cyanobacteria take up Fe are largely unknown, especially the initial step in Fe transport across the outer membrane. Here, we identified one TonB protein and four TonB-dependent transporters (TBDTs) of the energy-requiring Fe acquisition system and six porins of the passive diffusion Fe uptake system in the model cyanobacterium Synechocystis sp. PCC 6803. The results experimentally demonstrated that TBDTs not only participated in organic ferri-siderophore uptake, but also in inorganic, free Fe (Fe') acquisition. 55Fe uptake rate measurements showed that a TBDT quadruple mutant acquired Fe at a slower rate than the wild type and lost nearly all ability to take up ferri-siderophores, indicating that TBDTs are critical for siderophore uptake. However, the mutant retained the ability to take up Fe' at 42% of the wild type Fe' uptake rate, suggesting additional pathways of Fe' acquisition besides TBDTs, likely by porins. Mutations in four of the six porin-encoding genes produced a low- Fe-sensitive phenotype, while mutation in all six genes was lethal to cell survival. These diverse outer membrane Fe uptake pathways reflect cyanobacterial evolution and adaptation under a range of Fe regimes across aquatic systems. [ABSTRACT FROM AUTHOR]