Finally, we show that signaling output of CHARMS can be titrated by changing the number of effector binding sites on the monomer. 8/10

In collaboration with Randal Halfmann @stowersinstitute.bsky.social, we replaced the DF with a synthetic amyloid forming sequence (poly-TA). Increasing numbers of TA repeats led to a loss of FRAP recovery and an increase in signaling output, confirming the requirement for oligomer stability. 7/10

What are the biophysical properties at the core of helical filaments required for successful signalosome signaling? Using FRAP we show that one key property is the oligomer stability, as complexes with monomer turnover are unable to signal. 6/10

In one of the first experiments I did for this project, I showed that the oligomerizing DF domain can be replaced with a de novo designed helical filament (DHF) domain. @flobnow.bsky.social further showed that the DF domain can be replaced with a bacterial homolog predicted to form filaments. 4/10

CHARMS reconstituted signaling from IL-1Rs (and TLRs) in cells. We could now engineer this simplified single protein to define the properties required for signalosome function. 3/10

Protein filaments composed of death fold (DF) domains are recurring in immune signaling. IL-1Rs and TLRs signal through a three component signalosome activating TRAF6. We engineered CHARMS, a single component signalosome by fusing TRAF6 binding sites directly to MyD88. 2/10