The Fletcher Lab

Our paper describing the metabolic regulation of giant E3 ubiquitin ligase RNF213 by the well-known energy currency ATP, is out now in Nature Communications. Here we used an activity-based probe of cysteine E3 activity, with Cryo-EM and protein biochemistry, to dissect the basic requirements of RNF213 E3 activation by the small molecule ATP. By developing a method to deliver activity-based probes into live cells, we find that RNF213 activity is sensitive to intracellular ATP concentration changes that occur following interferon stimulation. This suggests that the antiviral state installed by interferons (typically defined by the profile of ‘interferon-stimulated genes’ upregulated by interferon) is also likely to be defined by protein activity changes that occur independently of protein abundance change.