⮝ Full datasets listing
PXD061783
PXD061783 is an original dataset announced via ProteomeXchange.
Dataset Summary
| Title | The non-catalytic epsilon DNA polymerase subunit is an NPF motif recognition protein |
| Description | Short linear motifs (SLiMs) in disordered protein regions direct numerous protein–protein interactions, yet most remain uncharacterized. The Asn-Pro-Phe (NPF) motif is a well-known EH-domain ligand implicated in endocytosis, but here we reveal that the non-catalytic subunit of human DNA polymerase epsilon (POLE2) also serves as a general NPF-motif receptor. Using a quantitative “native holdup” assay, we find that POLE2 selectively binds diverse NPF-containing peptides, including canonical EH-domain ligands (e.g., SYNJ1) and previously uncharacterized motifs. Biochemical measurements and mutational analysis show that NPF motifs interact with a shallow pocket near the POLE2 C-terminus, and AlphaFold predictions confirm key roles for Y513, E520, and S522 in motif coordination. Proteome-scale affinity screens identify NPF-containing nuclear proteins (e.g., WDHD1, DONSON, TTF2) that bind POLE2 with micromolar affinities, and their motif mutations abolish binding in cell extracts. Although POLE2 primarily tethers the catalytic POLE subunit to replication forks, these results indicate that it can also recruit various NPF-bearing partners involved in replication, DNA repair, and transcription regulation. Notably, NPF motifs optimized for EH-domain binding can still associate with POLE2, highlighting the inherent degeneracy of SLiM-mediated networks. Overall, these findings establish POLE2 as a central hub linking replication with other processes via broad NPF-motif recognition. |
| HostingRepository | MassIVE |
| AnnounceDate | 2025-12-15 |
| AnnouncementXML | Submission_2025-12-15_01:48:47.844.xml |
| DigitalObjectIdentifier | |
| ReviewLevel | Non peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Unsupported dataset by repository |
| PrimarySubmitter | Salla Keskitalo |
| SpeciesList | scientific name: Homo sapiens; |
| ModificationList | No PTMs are included in the dataset |
| Instrument | timsTOF Pro 2 |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|---|---|---|
| 0 | 2025-03-13 01:18:09 | ID requested | |
| ⏵ 1 | 2025-12-15 01:48:48 | announced |
Publication List
| no publication |
Keyword List
| submitter keyword: SLiM, NPF, Proteomics, Native holdup, nHU, motif, DatasetType:Proteomics, BioID |
Contact List
| Markku Varjosalo | |
|---|---|
| contact affiliation | University of Helsinki |
| contact email | markku.varjosalo@helsinki.fi |
| lab head | |
| Gergo Gogl | |
| contact affiliation | Inserm CRCN |
| contact email | gergo.gogl@univ-cotedazur.fr |
| lab head | |
| Salla Keskitalo | |
| contact affiliation | University of Helsinki |
| contact email | salla.keskitalo@helsinki.fi |
| dataset submitter | |
Full Dataset Link List
| MassIVE dataset URI |
| Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://massive-ftp.ucsd.edu/v09/MSV000097307/ |
