RT Journal Article
SR Electronic
T1 High-throughput identification of dominant negative polypeptides in yeast
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 418608
DO 10.1101/418608
A1 Michael W. Dorrity
A1 Christine Queitsch
A1 Stanley Fields
YR 2018
UL http://biorxiv.org/content/early/2018/09/15/418608.abstract
AB Dominant negative polypeptides can act as inhibitors by binding to the wild type protein or by titrating an essential ligand. Here, we use high-throughput sequencing of DNA libraries composed of fragments of yeast genes to identify dominant negative polypeptides based on their depletion during cell growth. The method uncovers numerous inhibitory polypeptides for a protein and thus is capable of defining interacting domains with exquisite resolution, even pinpointing individual residues that contact ligands.