Dataset Information

Therapeutic targeting of RNA splicing catalysis through inhibition of protein arginine methylation

ABSTRACT: Cancer-associated mutations in RNA splicing factors commonly occur in myeloid and lymphoid leukemias as well as a variety of solid tumors and confer dependence on wild-type (WT) splicing. These observations have led to clinical efforts to directly inhibit the spliceosome in patients with refractory leukemias. Here we identify that inhibiting symmetric (SDMA) or asymmetric dimethyl arginine methylation (ADMA), mediated by PRMT5 and type I protein arginine methyltransferases (PRMTs) respectively, reduces splicing fidelity and results in strong preferential killing of spliceosomal mutant leukemias over WT counterparts. Consistent with this, proteomic analyses identified RNA binding proteins and splicing factors as the most enriched PRMT5 and/or PRMT Type I substrates in leukemia. Accordingly, combined PRMT5/type I PRMT inhibition resulted in synergistic cell killing and pronounced effects on splicing compared with inhibiting either enzymatic activity alone. These data identify genetic subsets of cancer most likely to respond to PRMT inhibition and a mechanistic basis for the therapeutic efficacy of PRMT inhibition in cancer.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Bone Marrow, Blood

DISEASE(S): Acute Leukemia

SUBMITTER: Enrico Massignani

LAB HEAD: Tiziana Bonaldi

PROVIDER: PXD012007 | Pride | 2019-07-30

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	HF170710_DM_SGC_NB4_GSK591_FWD_SILAC_input_inSol_01.raw	Raw
	HF170710_DM_SGC_NB4_GSK591_FWD_SILAC_input_inSol_02.raw	Raw
	HF170710_DM_SGC_NB4_GSK591_FWD_SILAC_input_inSol_03.raw	Raw
	HF170710_DM_SGC_NB4_GSK591_FWD_SILAC_input_inSol_04.raw	Raw
	HF170710_DM_SGC_NB4_GSK591_FWD_SILAC_input_inSol_05.raw	Raw

Items per page:

1 - 5 of 189

Publications

Therapeutic Targeting of RNA Splicing Catalysis through Inhibition of Protein Arginine Methylation.

Fong Jia Yi JY Pignata Luca L Goy Pierre-Alexis PA Kawabata Kimihito Cojin KC Lee Stanley Chun-Wei SC Koh Cheryl M CM Musiani Daniele D Massignani Enrico E Kotini Andriana G AG Penson Alex A Wun Cheng Mun CM Shen Yudao Y Schwarz Megan M Low Diana Hp DH Rialdi Alexander A Ki Michelle M Wollmann Heike H Mzoughi Slim S Gay Florence F Thompson Christine C Hart Timothy T Barbash Olena O Luciani Genna M GM Szewczyk Magdalena M MM Wouters Bas J BJ Delwel Ruud R Papapetrou Eirini P EP Barsyte-Lovejoy Dalia D Arrowsmith Cheryl H CH Minden Mark D MD Jin Jian J Melnick Ari A Bonaldi Tiziana T Abdel-Wahab Omar O Guccione Ernesto E

Cancer cell 20190801 2

Cancer-associated mutations in genes encoding RNA splicing factors (SFs) commonly occur in leukemias, as well as in a variety of solid tumors, and confer dependence on wild-type splicing. These observations have led to clinical efforts to directly inhibit the spliceosome in patients with refractory leukemias. Here, we identify that inhibiting symmetric or asymmetric dimethylation of arginine, mediated by PRMT5 and type I protein arginine methyltransferases (PRMTs), respectively, reduces splicing ...[more]

PMID: 31408619

Similar Datasets

Project description:PRMT5 is the major enzyme that catalyzes the symmetric dimethylation of arginine (sDMA) in mammalian cells. Its activity is crucial for many biological processes including transcriptional regulation, mRNA processing, as well as DNA damage and repair. However, the protein substrates identified for PRMT5 have yet been limited in numbers, hindering the understanding of PRMT5 functions in normal as well as diseased cells. Herein we developed an optimized strategy for proteomic profiling of cellular sDMA and apply it to the discovery of novel PRMT5 substrates. Our results show that a combination of proteolytic digestion by the metalloprotease ulilysin and using electron-transfer dissociation with supplemental activation as the mass spectrometry method significantly increased sDMA site identification and localization after immuno-affinity enrichment. By employing SILAC-based differential quantitative proteomic approach and a PRMT5 specific inhibitor, we identified 325 sDMA sites being regulated by PRMT5, 220 being novel sites, which greatly expanded the number of PRMT5 substrates. Biochemical studies confirmed the newly discovered PRMT5 substrate, Plasminogen activator inhibitor 1 RNA-binding protein (SERBP1) and revealed that the RGG/RG motif in the central region of SERBP1 contains both PRMT5-catalyzed sDMA and Type I PRMT-catalyzed asymmetric dimethylation of arginine (aDMA). Unexpectedly, using the optimized methylarginine profiling strategy, we also discovered arginine trimethylation, a previously unknown type of modification, on the central RGG/RG region of SERBP1. By mutational studies we demonstrated that the trimethyl modification on R172 of SERBP1 regulates its recruitment to stress granule (SG) under oxidative stress, indicating a functional role of arginine trimethylation in the cell. Overall, the optimized profiling strategy not only enabled the identification of extensive, non-overlapping sDMA sites and novel PRMT5 substrates, but also revealed a potentially important new PTM, arginine trimethylation. The work lays the foundation for further investigations on the functional interplay of different methylation modifications on arginines, especially in the heavily methylated RGG/RG motif of many RNA-binding proteins.