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Large-scale map of RNA-binding protein interactomes across the mRNA life cycle.

Molecular cell · 2024 · Vol. 84 (19) · pp. 3790-3809.e8

Abstract

mRNAs interact with RNA-binding proteins (RBPs) throughout their processing and maturation. While efforts have assigned RBPs to RNA substrates, less exploration has leveraged protein-protein interactions (PPIs) to study proteins in mRNA life-cycle stages. We generated an RNA-aware, RBP-centric PPI map across the mRNA life cycle in human cells by immunopurification-mass spectrometry (IP-MS) of ∼100 endogenous RBPs with and without RNase, augmented by size exclusion chromatography-mass spectrometry (SEC-MS). We identify 8,742 known and 20,802 unreported interactions between 1,125 proteins and determine that 73% of the IP-MS-identified interactions are RNA regulated. Our interactome links many proteins, some with unknown functions, to specific mRNA life-cycle stages, with nearly half associated with multiple stages. We demonstrate the value of this resource by characterizing the splicing and export functions of enhancer of rudimentary homolog (ERH), and by showing that small nuclear ribonucleoprotein U5 subunit 200 (SNRNP200) interacts with stress granule proteins and binds cytoplasmic RNA differently during stress.

Publication Types

["Journal Article"]

Keywords

MeSH Terms

["Humans", "RNA, Messenger", "RNA-Binding Proteins", "Protein Interaction Maps", "Protein Binding", "HeLa Cells", "Protein Interaction Mapping", "Ribonucleoproteins, Small Nuclear", "HEK293 Cells", "Mass Spectrometry", "RNA Splicing"]

Funding

U24 HG009889 NHGRI NIH HHS (United States)
R01 HG004659 NHGRI NIH HHS (United States)
RF1 MH126719 NIMH NIH HHS (United States)
R01 HG011864 NHGRI NIH HHS (United States)
T32 CA067754 NCI NIH HHS (United States)
R01 HG012216 NHGRI NIH HHS (United States)
R35 GM128802 NIGMS NIH HHS (United States)
R01 AG071869 NIA NIH HHS (United States)
R01 CA284315 NCI NIH HHS (United States)
K22 NS112678 NINDS NIH HHS (United States)
K99 NS135103 NINDS NIH HHS (United States)

Linked Datasets (1)

GSE230717 GSE via ncbi_elink
GEO

Large-scale map of RNA binding protein interactomes across the mRNA life-cycle

Homo sapiens
48 data files
FileTypeSize
CAPRIN_US_IN1_S164_L001_R1_001.fastq.gz RIP-Seq 445.5 MB
CAPRIN_US_IN1_S164_L001_R1_001.fastq.gz RIP-Seq 445.5 MB
CAPRIN_US_IN2_S165_L001_R1_001.fastq.gz RIP-Seq 552.8 MB
CAPRIN_US_IN2_S165_L001_R1_001.fastq.gz RIP-Seq 552.8 MB
CAPRIN_US_IP1_S172_L001_R1_001.fastq.gz RIP-Seq 558.7 MB
CAPRIN_US_IP1_S172_L001_R1_001.fastq.gz RIP-Seq 558.7 MB
CAPRIN_US_IP2_S173_L001_R1_001.fastq.gz RIP-Seq 581.9 MB
CAPRIN_US_IP2_S173_L001_R1_001.fastq.gz RIP-Seq 581.9 MB
HEK-baseline-SNRNP200-Input_S22_L002_R1_001.fastq.gz RIP-Seq 930.2 MB
HEK-baseline-SNRNP200-Input_S22_L002_R1_001.fastq.gz RIP-Seq 930.2 MB
HEK-baseline-SNRNP200-IP_S23_L002_R1_001.fastq.gz RIP-Seq 1.0 GB
HEK-baseline-SNRNP200-IP_S23_L002_R1_001.fastq.gz RIP-Seq 1.0 GB
HEK-Input-SNRNP200_S15_L001_R1_001.fastq.gz RIP-Seq 575.7 MB
HEK-Input-SNRNP200_S15_L001_R1_001.fastq.gz RIP-Seq 575.7 MB
HEK-IP-SNRNP200_S16_L001_R1_001.fastq.gz RIP-Seq 535.0 MB
HEK-IP-SNRNP200_S16_L001_R1_001.fastq.gz RIP-Seq 535.0 MB
HEK-Stress-Input-SNRNP200_S3_L001_R1_001.fastq.gz RIP-Seq 523.6 MB
HEK-Stress-Input-SNRNP200_S3_L001_R1_001.fastq.gz RIP-Seq 523.6 MB
HEK-Stress-IP-SNRNP200_S4_L001_R1_001.fastq.gz RIP-Seq 642.9 MB
HEK-Stress-IP-SNRNP200_S4_L001_R1_001.fastq.gz RIP-Seq 642.9 MB
HEK-stress-SNRNP200-Input_S26_L002_R1_001.fastq.gz RIP-Seq 923.0 MB
HEK-stress-SNRNP200-Input_S26_L002_R1_001.fastq.gz RIP-Seq 923.0 MB
HEK-stress-SNRNP200-IP_S27_L002_R1_001.fastq.gz RIP-Seq 831.6 MB
HEK-stress-SNRNP200-IP_S27_L002_R1_001.fastq.gz RIP-Seq 831.6 MB
KR-A-IN_S50_L002_R1_001.fastq.gz RIP-Seq 354.2 MB
KR-A-IN_S50_L002_R1_001.fastq.gz RIP-Seq 354.2 MB
KR-A-IP_S49_L002_R1_001.fastq.gz RIP-Seq 143.5 MB
KR-A-IP_S49_L002_R1_001.fastq.gz RIP-Seq 143.5 MB
KR-B-IN_S52_L002_R1_001.fastq.gz RIP-Seq 306.5 MB
KR-B-IN_S52_L002_R1_001.fastq.gz RIP-Seq 306.5 MB
KR-B-IP_S51_L002_R1_001.fastq.gz RIP-Seq 293.1 MB
KR-B-IP_S51_L002_R1_001.fastq.gz RIP-Seq 293.1 MB
SNRNP200_cytoplasmic_arsenite_in1_S156_L001_R1_001.fastq.gz RIP-Seq 879.6 MB
SNRNP200_cytoplasmic_arsenite_in1_S156_L001_R1_001.fastq.gz RIP-Seq 879.6 MB
SNRNP200_cytoplasmic_arsenite_in2_S158_L001_R1_001.fastq.gz RIP-Seq 927.3 MB
SNRNP200_cytoplasmic_arsenite_in2_S158_L001_R1_001.fastq.gz RIP-Seq 927.3 MB
SNRNP200_cytoplasmic_arsenite_ip1_S157_L001_R1_001.fastq.gz RIP-Seq 965.0 MB
SNRNP200_cytoplasmic_arsenite_ip2_S159_L001_R1_001.fastq.gz RIP-Seq 1.0 GB
SNRNP200_cytoplasmic_control_in1_S152_L001_R1_001.fastq.gz RIP-Seq 673.7 MB
SNRNP200_cytoplasmic_control_in1_S152_L001_R1_001.fastq.gz RIP-Seq 673.7 MB
SNRNP200_cytoplasmic_control_in2_S154_L001_R1_001.fastq.gz RIP-Seq 662.6 MB
SNRNP200_cytoplasmic_control_in2_S154_L001_R1_001.fastq.gz RIP-Seq 662.6 MB
SNRNP200_cytoplasmic_control_ip1_S153_L001_R1_001.fastq.gz RIP-Seq 733.5 MB
SNRNP200_cytoplasmic_control_ip1_S153_L001_R1_001.fastq.gz RIP-Seq 733.5 MB
SNRNP200_cytoplasmic_control_ip2_S155_L001_R1_001.fastq.gz RIP-Seq 636.9 MB
SNRNP200_cytoplasmic_control_ip2_S155_L001_R1_001.fastq.gz RIP-Seq 636.9 MB
SRR24321506.lite RIP-Seq 1.0 GB
SRR24321507.lite RIP-Seq 965.0 MB

Potentially Related Datasets (2)

These accessions were text-mined from the PMC full text. They may be referenced for comparison, cited from other studies, or otherwise mentioned without being primary data for this paper.

Large-scale map of RNA binding protein interactomes across the mRNA life-cycle

MSV000093915 MASSIVE MassIVE

Analysis Pipelines (1)

eCLIP geo_data_processing GSE230717