Abstract 61: A genome-scale CRISPR-Cas9 screening method for protein stability reveals novel regulators of Cdc25A

Abstract

Abstract The regulation of stability is particularly crucial for unstable proteins in cells. However, a convenient and unbiased method of identifying regulators of protein stability remains to be generated. Recently, genome-scale CRISPR-Cas9 library has been established as a genetic tool to mediate loss of function screening. Here, we developed a method named Protein Stability Regulators Screening Assay (Pro-SRSA) by combining the whole genome CRISPR-Cas9 library with a dual-fluorescence-based protein stability reporter and high-throughput sequencing to screen regulators of protein stability. Using Cdc25A as an example, Cul4B-DDB1-DCAF8 was determined to be a new E3 ligase for Cdc25A. Moreover, the acetylation of Cdc25A at lysine 150, which was mediated by p300/CBP and deacetylated by HDAC3, was revealed to prevent its ubiquitin-mediated degradation by proteasome. This is the first report that the acetylation as a novel posttranslational modification modulates the Cdc25A stability, and we believe that this efficient and unbiased screening method at the genome-scale will be widely used to globally identify regulators of protein stability. Citation Format: Yuanzhong Wu, Tiebang Kang. A genome-scale CRISPR-Cas9 screening method for protein stability reveals novel regulators of Cdc25A. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 61.