PXD015196

PXD015196 is an original dataset announced via ProteomeXchange.

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

Revision	Datetime	Status	ChangeLog Entry
0	2019-08-28 07:04:41	ID requested
⏵ 1	2020-01-13 04:37:59	announced

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]

curator keyword: Biomedical

submitter keyword: Target identification, LC-MS, AfBPP, Thermal proteome profiling, co-IP, natural product

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

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://ftp.pride.ebi.ac.uk/pride/data/archive/2020/01/PXD015196

PRIDE project URI

• PRIDE
  1. PXD015196
     1. Label: PRIDE project
     2. Name: The cytotoxic natural product vioprolide A targets nucleolar protein 14 essential for ribosome biogenesis