14-3-3 Proteins Restrain the Exo1 Nuclease to Prevent Over-resection [Cell Biology]

April 1st, 2015 by Chen, X., Kim, I.-K., Honaker, Y., Paudyal, S. C., Koh, W. K., Sparks, M., Li, S., Piwnica-Worms, H., Ellenberger, T., You, Z.

The DNA end resection process dictates the cellular response to DNA double-strand break damage and is essential for genome maintenance. While insufficient DNA resection hinders homology-directed repair and the ATR-dependent checkpoint activation, over-resection produces excessive single-stranded DNA that could lead to genomic instability. However, how DNA end resection is properly controlled remains poorly understood. Here we show that the major resection nuclease Exo1 is regulated both positively and negatively by protein-protein interactions to ensure a proper level of DNA resection. We have previously shown that the sliding DNA clamp PCNA associates with the C-terminal domain of Exo1 and promotes Exo1 damage association and DNA resection. In this report we show that 14-3-3 proteins interact with a central region of Exo1 and negatively regulate Exo1 damage recruitment and subsequent resection. 14-3-3s limit Exo1 damage association, at least in part, by suppressing its association with PCNA. Disruption of Exo1-14-3-3 interaction results in elevated sensitivity of cells to DNA damage. Unlike Exo1, the Dna2 resection pathway is apparently not regulated by PCNA and 14-3-3s. Our results provide critical insights into the mechanism and regulation of the DNA end resection process and may have implications for cancer treatment.