PXD015196 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | The cytotoxic natural product vioprolide A targets nucleolar protein 14 essential for ribosome biogenesis |
Description | The cytotoxic and antifungal peptolides vioprolide A-D were discovered in 1996, however their mode-of-action has so far been unsolved. Anti-cancer effect of the most potent cytotoxic derivative vioprolide A (VioA) were evaluated, showing a prominent potency against human acute lymphoblastic leukemia (ALL) cells. In order to decipher the molecular mode-of-action, a VioA-derived photoprobe (VioA-P) was synthesized and utilized for MS-based AfBPP experiments. In addition thermal protein profiling (TPP) experiments with unmodified VioA were conducted for target discovery. TPP experiments revealed a strong thermal stabilization of two proteins after filtering. NOP14, the protein showing the strongest stabilization effect could further be verified as target with continuative biochemical experiments. As various protein-protein interactions with NOP14 are essential in ribosome biogenesis, e.g. with NOC4L and EMG1 for proper maturation of 18S rRNA and 40S assembly and export, the interactome of NOP14 was revealed by MS-based co-Immunoprecipitation. Our findings indicate, that VioA treatment leads to a loss of interaction between NOP14 and EMG1, whereas the NOP14-NOC4L interface remained unperturbed. These findings could be verified independently by western blot, also showing that overall protein expression of NOP14, EMG1 and NOC4L remained unaltered upon VioA treatment. Delocalization of EMG1 from the nucleus into the cytoplasm as consequence of missing interaction with NOP14 could be verified by immunostaining. |
HostingRepository | PRIDE |
AnnounceDate | 2020-01-13 |
AnnouncementXML | Submission_2020-01-13_04:37:57.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Volker Kirsch |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | No PTMs are included in the dataset |
Instrument | Orbitrap Fusion; Q Exactive |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2019-08-28 07:04:41 | ID requested | |
⏵ 1 | 2020-01-13 04:37:59 | announced | |
Publication List
Kirsch VC, Orgler C, Braig S, Jeremias I, Auerbach D, M, ü, ller R, Vollmar AM, Sieber SA, A Targets Nucleolar Protein 14, Which Is Essential for Ribosome Biogenesis. Angew Chem Int Ed Engl, 59(4):1595-1600(2020) [pubmed] |
Keyword List
curator keyword: Biomedical |
submitter keyword: Target identification, LC-MS, AfBPP, Thermal proteome profiling, co-IP, natural product |
Contact List
Stephan A. Sieber |
contact affiliation | Stephan A. Sieber, Center for Integrated Protein Science at the Department of Chemistry, Technische Universitä¤t München, Garching b. München 85747, Germany; Chair of Organic Chemistry II, Technische Universität München, Garching b. München 85747, Germany |
contact email | stephan.sieber@tum.de |
lab head | |
Volker Kirsch |
contact affiliation | Technical University of Munich |
contact email | volker.kirsch@tum.de |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
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[ - ]
- PRIDE
- PXD015196
- Label: PRIDE project
- Name: The cytotoxic natural product vioprolide A targets nucleolar protein 14 essential for ribosome biogenesis