RT @hattaca: Excited to share our lab's first preprint: we used spatial transcriptomics to dissect how aging disrupts the cycling ovary! 🧵….

RT @BingxuL: We are all born with a genetic lottery. Millions of T cell receptors are what we have with a hope to defend all cancer and vir….

RT @gheimberg: 1/ Introducing SCimilarity, a new foundation model to explore scRNAseq data across tissues and diseases! It learns a common….

Overall, we are excited for this approach as a way to interrogate sequence-function relationships for all types of proteins with complex activities throughout the cell. Please see our preprint for the full story! (14/14).

This requirement for a precise physical organization of LAT and interacting molecules may enforce downstream pathway balance, and we are curious to see whether similar receptor-adapter systems involving disordered proteins engage similar mechanisms. (13/14).

In short, we found that mutation of one position indirectly disrupts protein binding at distant sites. This is likely due to the dual role of these proteins as effectors of downstream pathways and bridging factors between LAT molecules. (12/14)

We then used protein proximity labeling with TurboID to ask how the assembly of LAT and its interacting proteins changes when we mutate individual positions in LAT. (11/14)

Why might this be the case? We relied on previous efforts characterizing LAT and AlphaFold-Multimer to zero in on a small number of direct LAT-binding proteins. (10/14)

Another major goal was to understand how each segment of LAT contributes to each of the downstream signaling pathways. This analysis led us to a surprising result – LAT mutants generally confer coordinated defects, balanced across all downstream pathways. (9/14)

By mutating several positions at a time to alter the local charge in LAT, we found that this feature is indeed important for LAT function. (8/14)

The other cool thing about disordered proteins is that their distributed biophysical features can influence function. The CIDER tool pointed to negative charge, a feature conserved in LAT even across species that have low conservation of precise amino acid sequence. (7/14)

How do these functional segments contribute to T cell activation? We first annotated short, conserved protein sequence motifs, which are commonly found throughout intrinsically disordered proteins and often serve as binding sites for partner proteins. (6/14)

At the highest level, we found functional sequence throughout the length of LAT, including known critical positions and previously uncharacterized segments. (5/14)

We delivered these mutants to T cells and employed high-content screening (Perturb-seq and Spear-ATAC), allowing us to link the presence of a LAT mutant in a cell to a rich measure of signaling pathway activities. (4/14)

We designed a large set of mutants to LAT, with the goal of understanding how each position of the protein contributes to LAT’s function in T cell activation. (3/14)

In this project with Tyler Dao, Aviv Regev, and Alex Shalek (@shaleklab, @broadinstitute, @MIT), we were interested in how the adapter LAT, an intrinsically disordered protein, channels T cell receptor stimulation into a balanced collection of signaling pathways. (2/14)

Excited to share our study of T cell signaling, where we directed high-content, single-cell genetic screening to uncover how a single adapter protein orchestrates a complex cellular process essential for health. (1/14).

RT @hattaca: Excited to announce I will start as an Assistant Professor @Yale @YaleMed on Jan 1, 2024! Our lab's mission is to build tools….

RT @DSilverbush: 🧬👩‍🔬Exciting News🧪🔬.The Silverbush Lab is opening soon at UPenn! The lab will decipher tumor #heterogeneity and #plasticit….

RT @gheimberg: Ever analyze a scRNAseq dataset and wonder if a specific cell state has been seen before? And if so, where in the human body….