MassIVE MSV000089544

Partial Public

CRISPR screening identifies C1orf112 as a novel modulator of the response to ICLs

Description

DNA interstrand crosslinks (ICLs) are highly cytotoxic DNA lesions that are commonly induced by endogenous metabolic processes and chemotherapeutic drugs, such as cisplatin. To prevent detrimental effects associated with ICLs, such as interference with both transcription and replication, cells rely on a complex coordination of different DNA repair pathways, including Fanconi Anemia (FA) and homologous recombination (HR), for their resolution. To unveil the breadth of factors that plays a role in this response, we performed CRISPR-based genome-wide screens treated with the clinically relevant cyclophosphamide agent. This approach, along with a fluorescence-based competition assay, defined C1orf112 as a novel modulator of the response to ICLs-inducing agents. Subsequent analysis showed that depletion of C1orf112 impairs genomic stability by increasing spontaneous levels of gamma-H2AX, micronuclei, and 53BP1-nuclear bodies. Using immunofluorescence approaches, we found that C1orf112 acts downstream of the FA pathways and is required for resolving ICL-induced RAD51 foci. Consistently, our results show that C1orf112 is required for homology-directed DNA repair, including HR and single-strand annealing. Proximal mapping of C1orf112 using TurboID technology identified the AAA+ ATPase FIGNL1 as a constitutive interactor and functional characterization of this complex shows that these C1orf112 and FIGNL1 cooperate in the unloading of RAD51 at ICL-induced lesion. Altogether, our findings reveal C1orf112 as a previously unidentified regulator of the AAA+ ATPase FIGNL1 in the resolution of ICLs by homology-directed DNA repair pathways. [doi:10.25345/C5MP4VR92] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: DNA repair ; Homologous recombination ; Fanconi Anemia ; CRISPR ; DNA double-strand breaks ; Bioid ; proximity labeling ; C1orf112 ; FIGNL1

Contact

Principal Investigators:
(in alphabetical order)
Alexandre Orthwein, McGill University, Canada
Submitting User: Monod
Number of Files:
Total Size:
Spectra:
Subscribers:
 
Owner Reanalyses
Experimental Design
    Conditions:
    Biological Replicates:
    Technical Replicates:
 
Identification Results
    Proteins (Human, Remapped):
    Proteins (Reported):
    Peptides:
    Variant Peptides:
    PSMs:
 
Quantification Results
    Differential Proteins:
    Quantified Proteins:
 
Browse Dataset Files
 
FTP Download Link (click to copy):

- Dataset Reanalyses


+ Dataset History


Click here to queue conversion of this dataset's submitted spectrum files to open formats (e.g. mzML). This process may take some time.

When complete, the converted files will be available in the "ccms_peak" subdirectory of the dataset's FTP space (accessible via the "FTP Download" link to the right).
Number of distinct conditions across all analyses (original submission and reanalyses) associated with this dataset.

Distinct condition labels are counted across all files submitted in the "Metadata" category having a "Condition" column in this dataset.

"N/A" means no results of this type were submitted.
Number of distinct biological replicates across all analyses (original submission and reanalyses) associated with this dataset.

Distinct replicate labels are counted across all files submitted in the "Metadata" category having a "BioReplicate" or "Replicate" column in this dataset.

"N/A" means no results of this type were submitted.
Number of distinct technical replicates across all analyses (original submission and reanalyses) associated with this dataset.

The technical replicate count is defined as the maximum number of times any one distinct combination of condition and biological replicate was analyzed across all files submitted in the "Metadata" category. In the case of fractionated experiments, only the first fraction is considered.

"N/A" means no results of this type were submitted.
Originally identified proteins that were automatically remapped by MassIVE to proteins in the SwissProt human reference database.

"N/A" means no results of this type were submitted.
Number of distinct protein accessions reported across all analyses (original submission and reanalyses) associated with this dataset.

"N/A" means no results of this type were submitted.
Number of distinct unmodified peptide sequences reported across all analyses (original submission and reanalyses) associated with this dataset.

"N/A" means no results of this type were submitted.
Number of distinct peptide sequences (including modified variants or peptidoforms) reported across all analyses (original submission and reanalyses) associated with this dataset.

"N/A" means no results of this type were submitted.
Total number of peptide-spectrum matches (i.e. spectrum identifications) reported across all analyses (original submission and reanalyses) associated with this dataset.

"N/A" means no results of this type were submitted.
Number of distinct proteins quantified across all analyses (original submission and reanalyses) associated with this dataset.

Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

"N/A" means no results of this type were submitted.
Number of distinct proteins found to be differentially abundant in at least one comparison across all analyses (original submission and reanalyses) associated with this dataset.

A protein is differentially abundant if its change in abundance across conditions is found to be statistically significant with an adjusted p-value <= 0.05 and lists no issues associated with statistical tests for differential abundance.

Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

"N/A" means no results of this type were submitted.
This dataset may not contain all raw spectra data as originally deposited in PRIDE. It has been imported to MassIVE for reanalysis purposes, so its spectra data here may consist solely of processed peak lists suitable for reanalysis with most software.