DOI: 10.5061/dryad.34tmpg4tn
title: Data from: USP37 prevents premature disassembly of stressed replisomes by TRAIP

description: The E3 ubiquitin ligase TRAIP associates with the replisome and helps this
 molecular machine deal with replication stress. While TRAIP promotes DNA
 inter-strand crosslink repair by triggering the disassembly of
 CDC45-MCM2-7-GINS (CMG) helicases that have converged on these lesions,
 disassembly of single CMGs that have stalled temporarily would be
 deleterious, suggesting that TRAIP must be carefully regulated. Here, we
 demonstrate that human cells lacking the de-ubiquitylating
 enzyme USP37 are hypersensitive to topoisomerase poisons and
 other replication stress-inducing agents. We further show that TRAIP loss
 rescues the hypersensitivity of USP37 knockout cells to topoisomerase
 inhibitors. In Xenopus egg extracts depleted of USP37, TRAIP promotes
 premature CMG ubiquitylation and disassembly when replisomes stall before
 they have fully converged. Finally, we provide evidence that USP37 binds
 replisomes via CDC45, and that this interaction mediates USP37’s response
 to topological stress. Based on these results, we propose that USP37
 protects genome stability by preventing TRAIP-dependent CMG unloading when
 replication stress impedes timely termination.
description: For CRISPR/Cas9 screens, biological duplicates of U2OS Cas9
 wild-type and <em>USP37 </em>KO<em> </em>cells
 were transduced at a MOI of 0.3 and 500-fold coverage of the Brunello
 library (Doench et al., 2016). Afterward, transductants were selected with
 1.5 µg/mL puromycin for 12 days. Library preparation and next-generation
 sequencing of the samples was performed as described previously (Bowden et
 al., 2020). Guide-enrichment analysis was performed using DrugZ (Colic et
 al., 2019).
description: # USP37 prevents premature disassembly of stressed replisomes by TRAIP
 [https://doi.org/10.5061/dryad.34tmpg4tn](https://doi.org/10.5061/dryad.34tmpg4tn) #### Description of the data and file structure Counts and analyses of CRISPR screens. Suffixes indicate the type of data: * Fig*****_drugz_result.xlsx – Results from analyses using DrugZ. Comparisons were of DMSO vs camptothecin treated cells of the same genotype. See DrugZ paper for details of the algorithm and precise definitions of the statistics reported ([https://doi.org/10.1186/s13073-019-0665-3)](https://doi.org/10.1186/s13073-019-0665-3). Columns defined below: * GENE: Gene symbol, as used in the counts files. * sumZ: Un-normalised summed Z-scores per gene. * numObs: Number of observations (guides) per gene. * normZ: Normalised summed Z-score. * pval_synth: p-value of synthetic lethality (i.e. the combination of guides targeting the named gene and the treatment caused decreased abundance). * rank_synth: Rank when ordered by most synthetically lethal. * fdr_synth: False discovery rate corrected significance of pval_synth. * pval_supp: p-value of synthetic viability (the opposite of synthetic lethality). * rank_supp: Rank when ordered by most synthetically viable. * fdr_supp: False discovery rate corrected significance of pval_supp. * Fig***_guide_counts.tsv** – CRISPR guide abundances * The first two columns ("guide" and "gene") give CRISPR guide id and gene symbol for each row. Subsequent columns are guide abundances for the named sample. * sample details: * | Replicate | Treatment | Background | | | :------------------------------- | :----------- | :--------- | :- | | WT-Essentialome\_S4 | Essentialome | WT | | | WT-Initial | Initial | WT | | | USP37KO-Clone17-Essentialome\_S9 | Essentialome | USP37KO | | | USP37KO-Clone6-Essentialome\_S5 | Essentialome | USP37KO | | | USP37KO-Initial-B2\_S1 | Initial | USP37KO | | | USP37KO-Initial-B1\_S1 | Initial | USP37KO | | | WT2-CTRL\_S5 | CONTROL | WT | | | WT-CTRL\_S1 | CONTROL | WT | | | WT-CPT\_S2 | Camptothecin | WT | | | WT2-CPT\_S6 | Camptothecin | WT | |