With Matt Evans, Ethan Veit, Blake Richardson, et al. at Mount Sinai. Analysis by Caroline Kikawa.
STAT2 is an innate immune signaling protein that can be bound and signaled for degradation by flaviviral non-structural protein 5. Human STAT2 is vulnerable to this targeting by NS5, but murine (mouse) STAT2 is not.
A yeast-2-hybrid (Y2H) library of STAT2 was created by the Evans lab. This library was designed to contain mutations in the 198-209 amino acid region of STAT2, where are all possible combinations of the 8 amino acid differences between human and mouse STAT2 were created. With 2^8 possible combinations, this results in 256 unique sequences.
They selected for yeast that uptook expression plasmids with '2x' media. They also selected the same library with '4x+' media, which selected for expression plasmid uptake as well as binding by NS5. Here, I analyze the result of barcoded subamplicon sequencing these libraries.
For a summary of the results, see analysis_notebook.ipynb. For interactive graphs, see the linked GitHub pages https://jbloomlab.github.io/ZIKV_Y2H_198-209_combinatorial_library/.
The input data are in ./data/: