Dataset Information

Arginine methylation and ubiquitylation crosstalk controls DNA end-resection and homologous recombination repair

ABSTRACT: Cross-talk between distinct protein post-translational modifications is critical for an effective DNA damage response. Arginine methylation plays an important role in maintaining genome stability, however how this modification integrates with other enzymatic activities is largely unknown. Here, we identify the deubiquitylating enzyme USP11 as a previously uncharacterised PRMT1 substrate, and that methylation of USP11 promotes DNA end-resection and the repair of DNA double strand breaks by homologous recombination (HR). In addition, we show that PRMT1 is a ubiquitylated protein that it is targeted for deubiquitylation by USP11, and that USP11 regulates the ability of PRMT1 to bind to and methylate MRE11. Interestingly, USP11 methylation by PRMT1 is not required for other USP11 activities during HR, such as PALB2 deubiquitylation. Taken together, our findings reveal a specific role for USP11 during the early stages of DSB repair, which is mediated through its ability to regulate the activity of the PRMT1-MRE11 pathway.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hek-293t Cell, Kidney

SUBMITTER: Mark Skehel

LAB HEAD: Clare C Davies

PROVIDER: PXD028324 | Pride | 2021-11-02

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	AMG5307A10LC2_B2_AMG5307A10LC2-40-248610.raw	Raw
	AMG5307A11LC2_C2_AMG5307A11LC2-40-248611.raw	Raw
	AMG5307A12LC2_D2_AMG5307A12LC2-40-248612.raw	Raw
	AMG5307A13LC2_E2_AMG5307A13LC2-40-248613.raw	Raw
	AMG5307A14LC2_F2_AMG5307A14LC2-40-248614.raw	Raw

Items per page:

1 - 5 of 115

Similar Datasets

Project description:Protein methylation has been implicated in many important biological contexts including signaling, metabolism, and transcriptional control. Despite the importance of this post- translational modification, the global analysis of protein methylation by mass spectrometry-based proteomics has not been extensively studied due to the lack of robust, well-characterized techniques for methyl-peptidemethyl peptide enrichment. Here, to better investigate protein methylation, we optimized and compared two methods for methyl-peptidemethyl peptide enrichment: immunoaffinity purification (IAP) and high pH strong cation exchange (SCX). Comparison of these methods revealed that they are largely orthogonal for monomethyl arginine (MMA), suggesting that the usage of both techniques is required to provide a global view of protein methylation. Using both IAP and SCX, we investigated changes in protein methylation downstream of protein arginine methyltransferase 1 (PRMT1). Together, these techniques allowed us to quantify over ~1,000 arginine methylation sites on 407 proteins. Of these methylation sites, PRMT1 knockdown resulted in significant changes to 110 arginine methylation sites on 59 proteins. In contrast, zero lysine methylation sites were significantly changed upon PRMT1 knockdown. In PRMT1 knockdown cells, 24 MMA sites exhibited both significant downregulation (ie, putative PRMT1-mediated MMA targets) and 63 significant upregulation (ie, putative PRMT1-mediated ADMA targets). PRMT1 knockdown induced significant changes in asymmetric dimethyl arginine (ADMA), consistent with being PRMT1 targets. Additionally, We also observed that PRMT1 knockdown induced significant increases in symmetric dimethyl arginine (SDMA) methylation, suggesting that loss of PRMT1 activity allows scavenging of PRMT1 substrates by Type II PRMTs. Analysis of the subcellular localization and protein function of the significantly changing methyl-proteins revealed that the majority of PRMT1 substrates are localized to the nucleus and involved in RNA and DNA binding. Taken together, our results suggest that deep protein methylation profiling by mass spectrometry requires orthogonal enrichment techniques, identify novel PRMT1 methylation targets, and highlight in order to understand the dynamic interplay between methyltransferases in mammalian cells.

Dataset Information

Arginine methylation and ubiquitylation crosstalk controls DNA end-resection and homologous recombination repair

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets