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bioRxiv · 10.1101/2025.10.07.680950

The 9-1-1 complex protects ssDNA gaps in BRCA2-deficient cancer

Grimsley, H. E.·Courtemanche, K.·Cox, S.·McDermott, N.·Sharma, A.·Bright, J.·Setton, J.·Orman, M.·Powell, S. N. N.
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Abstract

Single-stranded DNA (ssDNA) gaps are a hallmark of BRCA-deficient cells, yet the mechanisms that safeguard these lesions remain unclear. Through a genome-wide CRISPR screen, we identified the RAD9A-HUS1-RAD1 (9-1-1) complex as essential for the survival of BRCA2-deficient cells through an ATR-independent mechanism. Loss of 9-1-1 in this context leads to the accumulation of PRIMPOL-dependent gaps that fail to undergo post-replicat ive repair, resulting in pathological expansion and increased DNA damage. This instability is driven by excessive EXO1-mediated degradation, as EXO1 depletion rescues the phenotype. We further demonstrate that the 9-1-1 complex is required for POL{zeta}-dependent gap filling. We propose a model in which ssDNA gaps, when extended beyond a critical length, become inaccessible to TLS-mediated repair and are fully reliant on homologous recombination. These findings establish the 9-1-1 complex as key regulator of ssDNA gap stability and a promising therapeutic target in BRCA2-deficient cancers. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/680950v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@56516aorg.highwire.dtl.DTLVardef@fb1ce3org.highwire.dtl.DTLVardef@25a7b2org.highwire.dtl.DTLVardef@249731_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Grimsley, H. E., Courtemanche, K., Cox, S., McDermott, N., Sharma, A., Bright, J., Setton, J., Orman, M., Powell, S. N. N.. 2025-10-07. The 9-1-1 complex protects ssDNA gaps in BRCA2-deficient cancer. https://doi.org/10.1101/2025.10.07.680950

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