Hijacking the host proteasome for the temporal degradation of bacterial effectors

To establish infection, intracellular pathogens need to modulate host cellular processes. Modulation of host processes is achieved by the action of various "effector proteins" which are delivered from the bacteria to the host cell cytosol. In order to orchestrate host cell reprogramming, the function of effectors inside host cells is regulated both temporally and spatially. In eukaryotes one of the most prominent processes used to degrade proteins is the ubiquitin-proteasome system. Recently it has emerged that the intracellular pathogen Legionella pneumophila is able to achieve temporal regulation of an effector using the ubiquitin-proteasome system. After establishing its replicative niche, the L. pneumophila effector SidH is degraded by the host proteasome. Most remarkably another effector protein LubX is able to mimic the function of an eukaryotic E3 ubiquitin ligase and polyubiquitinates SidH, targeting it for degradation. In this paper we describe a method to detect the polyubiquitin-modified forms of SidH in vitro and in vivo. Analyzing the temporal profile of polyubiquitination and degradation of bacterial effectors aids towards our understanding of how bacteria hijack host systems.