Abstract
DNA damage-activated signaling pathways are critical for coordinating multiple cellular processes, which must be tightly regulated to maintain genome stability. To provide a comprehensive and unbiased perspective of DNA damage response (DDR) signaling pathways, we performed 30 fluorescence-activated cell sorting (FACS)-based genome-wide CRISPR screens in human cell lines with antibodies recognizing distinct endogenous DNA damage signaling proteins to identify critical regulators involved in DDR. We discovered that proteasome-mediated processing is an early and prerequisite event for cells to trigger camptothecin- and etoposide-induced DDR signaling. Furthermore, we identified PRMT1 and PRMT5 as modulators that regulate ATM protein level. Moreover, we discovered that GNB1L is a key regulator of DDR signaling via its role as a co-chaperone specifically regulating PIKK proteins. Collectively, these screens offer a rich resource for further investigation of DDR, which may provide insight into strategies of targeting these DDR pathways to improve therapeutic outcomes.
Original language | English (US) |
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Pages (from-to) | 2810-2828.e6 |
Journal | Molecular cell |
Volume | 83 |
Issue number | 15 |
DOIs | |
State | Published - Aug 3 2023 |
Keywords
- antibody
- DDR signaling
- FACS-based CRISPR screen
- GNB1L
- PRMT5
- proteasome
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology