Proteomics

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Cisplatin induces the rearrangement of RBPs on CDKN1B/p27 mRNA

ABSTRACT: Cisplatin (CP) is a chemotherapeutic drug that is used to cure different types of cancer. CP induces DNA damage and leads to cell cycle arrest. The cyclin-dependent kinase inhibitor 1B (CDKN1B), also termed p27, plays an important role in the drug response ; and increased levels of p27 correlated with CP resistance. In HEK293 cells, we observed that p27 mRNAs levels increased whereas protein level drastically decreased in cells treated with CP; suggesting post-transcriptional regulatory events. To further understand the underlying mechanisms, we applied a biochemical approach combined with mass-spectrometry to systematically identify the RNA-binding proteins (RBPs) that are bound to the 3’UTR of p27 mRNAs in CP-treated versus non-treated cells in vivo. We found that 24 proteins, most of them known RBPs such HuR, hNRNPD, changed their association with p27 mRNA upon CP treatement. Furthermore, knock-down of a subset of the identified RBPs led to the inhibition of the CP-induced increase of p27 mRNA levels. In conclusion, these results highlight substantial rearrangement between RBPs and p27 mRNA upon CP treatment and corroborate the importance of post-transcriptional control in cellular drug response.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Kidney

SUBMITTER: Valentina Iadevaia  

LAB HEAD: Valentina Iadevaia

PROVIDER: PXD008498 | Pride | 2019-09-27

REPOSITORIES: Pride

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Dataset's files

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Action DRS
CPtreated_P2.msf Msf
CPtreated_P2.raw Raw
CPtreated_P4.msf Msf
CPtreated_P4.raw Raw
CPtreated_P4a.msf Msf
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Publications

Tandem RNA isolation reveals functional rearrangement of RNA-binding proteins on <i>CDKN1B/p27</i><sup>Kip1</sup> 3'UTRs in cisplatin treated cells.

Iadevaia Valentina V   Wouters Maikel D MD   Kanitz Alexander A   Matia-González Ana M AM   Laing Emma E EE   Gerber André P AP  

RNA biology 20190916 1

Post-transcriptional control of gene expression is mediated via RNA-binding proteins (RBPs) that interact with mRNAs in a combinatorial fashion. While recent global RNA interactome capture experiments expanded the repertoire of cellular RBPs quiet dramatically, little is known about the assembly of RBPs on particular mRNAs; and how these associations change and control the fate of the mRNA in drug-treatment conditions. Here we introduce a novel biochemical approach, termed tobramycin-based tande  ...[more]

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