1️⃣ @mmbronstein.bsky.social (@aithyra.bsky.social Scientific Director of AI) spoke with @mlichtenstein.bsky.social (@georgwinter.bsky.social's group) about the value of taking on ambitious projects & the key role of collaboration in science.

Thanks!

This was a hugely collaborative project - thanks to all involved. Grateful for the amazing co-first authors @fakuncao.bsky.social and @flobnow.bsky.social. Thanks for the great supervision and ideas Marcus and the invaluable contributions of Paulina, Daniel, Anna, Elke and Randal. 10/10

With this work we lay out a reductionist framework for engineering signalosomes, thereby defining the biophysical properties required for their function. Our results also raise the possibility for the design of synthetic and/or orthogonal signalosomes to program cellular decision making. 9/10

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

The ability of a bacterial DF homolog to form a functional signalosome indicates a conservation of oligomerization across the DF superfamily. It also demonstrates the potential for engineering mammalian immune signaling by repurposing bacterial anti-phage systems. 5/10

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

@mlichtenstein.bsky.social and I also used facilitated dissociation to make biosensors which are just as modular as those built previously with our LOCKR platform, but which respond 70 times faster.

Also, if you have ideas for how these switchable cytokines can modulate biological systems in new ways, we would love to discuss/collaborate! We envision these tools could open up a whole new way to explore the biology of transient cytokine signaling.