DEEPN as an Approach for Batch Processing of Yeast 2-Hybrid Interactions

作者：Pashkova Natasha; Peterson Tabitha A; Krishnamani Venkatramanan; Breheny Patrick; Stamnes Mark; Piper Robert C^*

来源：Cell Reports, 2016, 17(1): 303-315.

DOI：10.1016/j.celrep.2016.08.095

We adapted the yeast 2-hybrid assay to simultaneously uncover multiple transient protein interactions within a single screen by using a strategy termed DEEPN (dynamic enrichment for evaluation of protein networks). This approach incorporates high-throughput DNA sequencing and computation to follow competition among a plasmid population encoding interacting partners. To demonstrate the capacity of DEEPN, we identify a wide range of ubiq-uitin-binding proteins, including interactors that we verify biochemically. To demonstrate the specificity of DEEPN, we show that DEEPN allows simultaneous comparison of candidate interactors across multiple bait proteins, allowing differential interactions to be identified. This feature was used to identify interactors that distinguish between GTP- and GDP-bound conformations of Rab5.

• 出版日期2016-9-27