A comprehensive two-hybrid analysis to explore theL. pneumophilaeffector-effector interactome

Abstract

AbstractLegionella pneumophilauses over 300 translocated effector proteins to rewire host cells during infection and create a replicative niche for intracellular growth. To date, several studies have identifiedL. pneumophilaeffectors that indirectly and directly regulate the activity of other effectors, providing an additional layer of regulatory complexity. Amongst these are “metaeffectors” – a special class of effectors that regulate the activity of other effectors once inside the host. A defining feature of metaeffectors is direct, physical interaction with a target effector. Metaeffector identification to date has depended on phenotypes in heterologous systems and experimental serendipity. Using a multiplexed, recombinant-barcode-based yeast two-hybrid technology we screened for protein-protein interactions amongst allL. pneumophilaeffectors and several components of the Dot/Icm type IV secretion system (>167,000 protein combinations). Of the 52 protein interactions identified by this approach, 44 are novel protein interactions, including ten novel effector-effector interactions (doubling the number of known effector-effector interactions).