PXD012045 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Depletion of the RNA binding protein HNRNPD impairs homologous recombination by inhibiting DNA-end resection and inducing R-loop accumulation |
Description | DNA double strand break (DSB) repair through homologous recombination (HR) is crucial to maintain genome stability. DSB resection generates a single strand DNA intermediate, which is crucial for the HR process. We used a synthetic DNA structure, mimicking a resection intermediate, as a bait to identify proteins involved in this process. Among these, LC/MS analysis identified the RNA binding protein, HNRNPD. We found that HNRNPD was able to bind chromatin, although this binding occurred independently of DNA damage. However, upon damage, HNRNPD re-localized to γH2Ax foci and its silencing impaired CHK1 S345 phosphorylation and the DNA end resection process. Indeed, HNRNPD silencing reduced the ssDNA fraction upon camptothecin treatment and AsiSI-induced DSB resection and reduced RPA32 S4/8 phosphorylation. CRISPR/Cas9-mediated HNRNPD knockout impaired in vitro DNA resection and sensitized cells to camptothecin and olaparib treatment. We found that HNRNPD interacts with the heterogeneous nuclear ribonucleoprotein SAF-A previously associated with DNA damage repair. HNRNPD depletion resulted in an increased amount of RNA:DNA hybrids upon DNA damage . Both the expression of RNase H1 and RNA pol II inhibition recovered the ability to phosphorylate RPA32 S4/8 in HNRNPD knockout cells upon DNA damage, suggesting that RNA:DNA hybrid resolution likely rescues the defective DNA damage response of HNRNPD-depleted cells. |
HostingRepository | PRIDE |
AnnounceDate | 2019-02-26 |
AnnouncementXML | Submission_2019-03-05_07:27:46.xml |
DigitalObjectIdentifier | https://dx.doi.org/10.6019/PXD012045 |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Supported dataset by repository |
PrimarySubmitter | Claudia Landi |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | Carbamidomethyl |
Instrument | LCQ Classic |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2018-12-13 01:45:10 | ID requested | |
1 | 2019-02-26 05:18:23 | announced | |
⏵ 2 | 2019-03-05 07:27:48 | announced | Updated publication reference for PubMed record(s): 30799487. |
Publication List
Alfano L, Caporaso A, Altieri A, Dell'Aquila M, Landi C, Bini L, Pentimalli F, Giordano A, Depletion of the RNA binding protein HNRNPD impairs homologous recombination by inhibiting DNA-end resection and inducing R-loop accumulation. Nucleic Acids Res, 47(8):4068-4085(2019) [pubmed] |
Keyword List
submitter keyword: RNA binding proteins |
HNRNPD |
DNA damage |
DNA end resection |
RNA binding proteins |
R-loop |
Contact List
Luca Bini |
contact affiliation | Functional Proteomics Lab, Life Sciences Dept. University of Siena, Siena, Italy |
contact email | luca.bini@unisi.it |
lab head | |
Claudia Landi |
contact affiliation | University of Siena |
contact email | landi35@unisi.it |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD012045
- Label: PRIDE project
- Name: Depletion of the RNA binding protein HNRNPD impairs homologous recombination by inhibiting DNA-end resection and inducing R-loop accumulation