Dataset Information

The cytotoxic natural product vioprolide A targets nucleolar protein 14 essential for ribosome biogenesis

ABSTRACT: 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.

INSTRUMENT(S): Orbitrap Fusion, Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Suspension Culture, T Cell

DISEASE(S): Acute Leukemia

SUBMITTER: Volker Kirsch

LAB HEAD: Stephan A. Sieber

PROVIDER: PXD015196 | Pride | 2020-01-13

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20181015humanfastafile.fasta	Fasta
	20181015humanfastafileAfBPP.fasta	Fasta
	20181015humanfastafileCoIP.fasta	Fasta
	20190222_VCKB83_051.raw	Raw
	20190222_VCKB83_052.raw	Raw

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Publications

The Cytotoxic Natural Product Vioprolide A Targets Nucleolar Protein 14, Which Is Essential for Ribosome Biogenesis.

Kirsch Volker C VC Orgler Christina C Braig Simone S Jeremias Irmela I Auerbach David D Müller Rolf R Vollmar Angelika M AM Sieber Stephan A SA

Angewandte Chemie (International ed. in English) 20191212 4

Novel targets are needed for treatment of devastating diseases such as cancer. For decades, natural products have guided innovative therapies by addressing diverse pathways. Inspired by the potent cytotoxic bioactivity of myxobacterial vioprolides A-D, we performed in-depth studies on their mode of action. Based on its prominent potency against human acute lymphoblastic leukemia (ALL) cells, we conducted thermal proteome profiling (TPP) and deciphered the target proteins of the most active deriv ...[more]

PMID: 31658409

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