LnaB: a Legionella pneumophila activator of NF‐κB

Summary Legionella pneumophila possesses a large arsenal of type IV translocated substrates. Over 100 such proteins have been identified, but the functions of most are unknown. Previous studies have demonstrated that L. pneumophila activates NF-κB, a master transcriptional regulator of the mammalian innate immune response. Activation of NF-κB is dependent on the Legionella Icm/Dot type IV protein translocation system, consistent with the possibility that translocated bacterial proteins contribute to this response. To test this hypothesis, an expression library of 159 known and putative translocated substrates was created to evaluate whether ectopic production of a single L. pneumophila protein could activate NF-κB in mammalian cells. Expression of two of these proteins, LnaB (LegionellaNF-κB activator B) and LegK1, resulted in ∼150-fold induction of NF-κB activity in HEK293T cells, levels similar to the strong induction that occurs with ectopic expression of the known activator Nod1. LnaB is a substrate of the Icm/Dot system, and in the absence of this protein, a partial reduction of NF-κB activation in host cells occurs after challenge by post-exponential phase bacteria. These data indicate that LnaB is an Icm/Dot substrate that contributes to NF-κB activation during L. pneumophila infection in host cells.