Author:
Christenson Eric T.,Isaac Dervla T.,Yoshida Karin,Lipo Erion,Kim Jin-Sik,Ghirlando Rodolfo,Isberg Ralph R.,Banerjee Anirban
Abstract
Legionella pneumophilacauses a potentially fatal form of pneumonia by replicating within macrophages in theLegionella-containing vacuole (LCV). Bacterial survival and proliferation within the LCV rely on hundreds of secreted effector proteins comprising high functional redundancy. The vacuolar membrane-localized MavN, hypothesized to support iron transport, is unique among effectors because loss-of-function mutations result in severe intracellular growth defects. We show here an iron starvation response byL. pneumophilaafter infection of macrophages that was prematurely induced in the absence of MavN, consistent with MavN granting access to limiting cellular iron stores. MavN cysteine accessibilities to a membrane-impermeant label were determined during macrophage infections, revealing a topological pattern supporting multipass membrane transporter models. Mutations to several highly conserved residues that can take part in metal recognition and transport resulted in defective intracellular growth. Purified MavN and mutant derivatives were directly tested for transporter activity after heterologous purification and liposome reconstitution. Proteoliposomes harboring MavN exhibited robust transport of Fe2+, with the severity of defect of most mutants closely mimicking the magnitude of defects during intracellular growth. Surprisingly, MavN was equivalently proficient at transporting Fe2+, Mn2+, Co2+, or Zn2+. Consequently, flooding infected cells with either Mn2+or Zn2+allowed collaboration with iron to enhance intracellular growth ofL. pneumophilaΔmavNstrains, indicating a clear role for MavN in transporting each of these ions. These findings reveal that MavN is a transition-metal-ion transporter that plays a critical role in response to iron limitation duringLegionellainfection.
Funder
HHS | NIH | Eunice Kennedy Shriver National Institute of Child Health and Human Development
HHS | NIH | National Institute of Allergy and Infectious Diseases
Publisher
Proceedings of the National Academy of Sciences
Cited by
20 articles.
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