GSE86035
GSE GEOSONAR discovers RNA binding proteins from analysis of large-scale protein-protein interactomes.
Relations
Summary
RNA metabolism is controlled by an expanding yet incomplete catalog of RNA binding proteins (RBPs), many of which lack characterized RNA binding domains. Approaches to expand the RBP repertoire to discover non-canonical RBPs are currently needed. Here, HaloTag fusion pull-down of twelve nuclear and cytoplasmic RBPs followed by quantitative mass-spectrometry (MS) demonstrates that proteins interacting with multiple RBPs in an RNA-dependent manner are enriched for RBPs. This motivated SONAR, a computational approach that predicts RNA binding activity by analyzing large-scale affinity precipitation-MS protein-protein interactomes. Without relying on sequence or structure information, SONAR identifies 1923 human, 489 fly and 745 yeast RBPs, including over 100 human candidate RBPs that contain zinc finger domains. Enhanced CLIP confirms RNA binding activity and identifies transcriptome-wide RNA binding sites for SONAR-predicted RBPs, revealing unexpected RNA binding activity for disease-relevant proteins and DNA binding proteins.
Overall Design
eCLIP-seq was performed in biological replicate for AIFM1, NUMA1, RANGAP1, RNF219, VIM, ZNF184. Each sample has a size-matched input control for analysis
Analysis (32 steps)
View Data Processing- Takes output from raw files.
- Run to trim off both 5â and 3â adapters on both reads.
- Command: quality-cutoff 6 -m 18 -a NNNNNAGATCGGAAGAGCACACGTCTGAACTCCAGTCAC -g CTTCCGATCTACAAGTT -g CTTCCGATCTTGGTCCT -A AACTTGTAGATCGGA -A AGGACCAAGATCGGA -A ACTTGTAGATCGGAA -A GGACCAAGATCGGAA -A CTTGT AGATCGGAAG -A GACCAAGATCGGAAG -A TTGTAGATCGGAAGA -A ACCAAGATCGGAAGA -A TGTAGATCGGAAGAG -A CCAAGATCGGAAGAG -A GTAGATCGGAAGAGC -A CAAGATCGGAAGAGC -A TAGATCGGAAGAGCG -A AAGATCGGAAGAGCG -A AGATCGGAAGAGCGT -A GATCGGAAGAGCGTC -A ATCGGAAGAGCGTCG -A TCGGAAGAGCGTCGT -A CGGAAGAGCGTCGTG -A GGAAGAGCGTCGTGT -o /full/path/to/files/file_R1.C01.fastq.gz.adapterTrim.fastq.gz -p /full/path/to/files/file_R2.C01.fastq.gz.adapterTrim.fastq.gz /full/path/to/files/file_R1.C01.fastq.gz /full/path/to/files/file_R2.C01.fastq.gz > /full/path/to/files/file_R1.C01.fastq.gz.adapterTrim.metrics
- Takes output from cutadapt round 1.
- Run to trim off the 3â adapters on read 2, to control for double ligation events.
- Command: cutadapt -f fastq --match-read-wildcards --times 1 -e 0.1 -O 5 --quality-cutoff 6 -m 18 -A AACTTGTAGATCGGA -A AGGACCAAGATCGGA -A ACTTGTAGATCGGAA -A GGACCAAGATCGGAA -A CTTGTAGATCGGAAG -A GACCAAGATCGGAAG -A TTGTAGATCGGAAGA -A ACCAAGATCGGAAGA -A TGTAGATCGGAAGAG -A CCAAGATCGGAAGAG -A GTAGATCGGAAGAGC -A CAAGATCGGAAGAGC -A TAGATCGGAAGAGCG -A AAGATCGGAAGAGCG -A AGATCGGAAGAGCGT -A GATCGGAAGAGCGTC -A ATCGGAAGAGCGTCG -A TCGGAAGAGCGTCGT -A CGGAAGAGCGTCGTG -A GGAAGAGCGTCGTGT -o /full/path/to/files/file_R1.C01.fastq.gz.adapterTrim.round2.fastq.gz -p /full/path/to/files/file_R2.C01.fastq.gz.adapterTrim.round2.fastq.gz /full/path/to/files/file_R1.C01.fastq.gz.adapterTrim.fastq.gz /full/path/to/files/file_R2.C01.fastq.gz.adapterTrim.fastq.gz > /full/path/to/files/file_R1.C01.fastq.gz.adapterTrim.round2.metrics
- Takes output from cutadapt round 2.
- Maps to human specific version of RepBase used to remove repetitive elements, helps control for spurious artifacts from rRNA (& other) repetitive reads.
Supplementary Files (1)
Dataset Citations (1)
SRA Experiments (17) and Runs (28)
Total: 9835 MBSample attributes
Original files (1)
Sample attributes
Original files (1)
Sample attributes
Original files (1)
Runs (1)
| Run | Spots | Bases | Size (MB) | Files | Link |
|---|---|---|---|---|---|
| SRR4063769 | 17865347 | 1875861435 | 656.71 | KB5_293_CLIP_RNF219_1_input_ATTACTCG-AGGCGAAG_L008_R1.unassigned.rand… | SRA |
Sample attributes
Original files (1)
Sample attributes
Original files (1)
Sample attributes
Original files (1)
Runs (1)
| Run | Spots | Bases | Size (MB) | Files | Link |
|---|---|---|---|---|---|
| SRR4063774 | 19396533 | 2036635965 | 704.33 | KB8_293_CLIP_RANGAP_2_input_ATTCAGAA-CCTATCCT_L008_R1.unassigned.rand… | SRA |
Sample attributes
Original files (1)
Sample attributes
Original files (1)
Sample attributes
Original files (1)
Runs (1)
| Run | Spots | Bases | Size (MB) | Files | Link |
|---|---|---|---|---|---|
| SRR4063779 | 17854574 | 1874730270 | 652.7 | KB11_293_CLIP_NUMA_1_input_TCCGGAGA-AGGCGAAG_L008_R1.unassigned.rando… | SRA |
Sample attributes
Original files (1)
Runs (1)
| Run | Spots | Bases | Size (MB) | Files | Link |
|---|---|---|---|---|---|
| SRR4063780 | 10276666 | 1079049930 | 377.3 | KB12_293_CLIP_AIFM_1_IP_ATTACTCG-CCTATCCT_L006_R1.unassigned.randomer… | SRA |
Sample attributes
Original files (1)
Sample attributes
Original files (1)
Sample attributes
Original files (1)
Sample attributes
Original files (1)
Runs (1)
| Run | Spots | Bases | Size (MB) | Files | Link |
|---|---|---|---|---|---|
| SRR4063787 | 17093709 | 1794839445 | 682.35 | KB22_293_CLIP_VIM_input_CGCTCATT-TAATCTTA_L007_R1.unassigned.randomer… | SRA |
Sample attributes
Original files (1)
Runs (1)
| Run | Spots | Bases | Size (MB) | Files | Link |
|---|---|---|---|---|---|
| SRR4063788 | 31801837 | 3816220440 | 1300.01 | KB23_293_INPUT_ZNF184_S70_L008_R1_001.unassigned.randomer.fastq.gz, K… | SRA |
Sample attributes
Original files (1)
Sample attributes
Original files (1)
Linked Publications (1)
Data Files (32)
| Accession | File Name | Stored Type | Output Type | Mapping Assembly | Size | Download | |
|---|---|---|---|---|---|---|---|
| — | KB10_293_CLIP_NUMA_2_IP_ATTACTCG-CCTATCCT_L008_R1.A01_… | RIP-Seq | 108.5 MB | link | |||
| — | KB10_293_CLIP_NUMA_2_IP_ATTACTCG-CCTATCCT_L008_R1.B06_… | RIP-Seq | 191.8 MB | link | |||
| — | KB10_293_CLIP_NUMA_2_IP_ATTACTCG-CCTATCCT_L008_R2.B06_… | RIP-Seq | 191.8 MB | link | |||
| — | KB11_293_CLIP_NUMA_1_input_TCCGGAGA-AGGCGAAG_L008_R1.u… | RIP-Seq | 652.7 MB | link | |||
| — | KB12_293_CLIP_AIFM_1_IP_ATTACTCG-CCTATCCT_L006_R1.unas… | RIP-Seq | 377.3 MB | link | |||
| — | KB16_293_CLIP_RNF219_3_GAATTCGT-AGGCGAAG_L007_R1.C01_K… | RIP-Seq | 136.2 MB | link | |||
| — | KB16_293_CLIP_RNF219_3_GAATTCGT-AGGCGAAG_L007_R1.D08fi… | RIP-Seq | 178.3 MB | link | |||
| — | KB20_293_CLIP_VIM_1_GAATTCGT-ATAGAGGC_L007_R1.C01_KB20… | RIP-Seq | 215.8 MB | link | |||
| — | KB20_293_CLIP_VIM_1_GAATTCGT-ATAGAGGC_L007_R1.D08fixed… | RIP-Seq | 89.3 MB | link | |||
| — | KB21_293_CLIP_VIM_2_CGCTCATT-ATAGAGGC_L007_R1.C01_KB21… | RIP-Seq | 256.4 MB | link | |||
| — | KB21_293_CLIP_VIM_2_CGCTCATT-ATAGAGGC_L007_R1.D08fixed… | RIP-Seq | 112.0 MB | link | |||
| — | KB22_293_CLIP_VIM_input_CGCTCATT-TAATCTTA_L007_R1.unas… | RIP-Seq | 682.3 MB | link | |||
| — | KB2_293_CLIP_AIFM_1_IP_TCCGGAGA-TATAGCCT_L008_R1.A01_K… | RIP-Seq | 131.6 MB | link | |||
| — | KB2_293_CLIP_AIFM_1_IP_TCCGGAGA-TATAGCCT_L008_R1.B06_K… | RIP-Seq | 130.8 MB | link | |||
| — | KB23_293_INPUT_ZNF184_S70_L008_R1_001.unassigned.rando… | RIP-Seq | 1.3 GB | link | |||
| — | KB24_293_CLIP_ZNF184_S71_L008_R1_001.A01_KB24_ZNF184_1… | RIP-Seq | 433.9 MB | link | |||
| — | KB24_293_CLIP_ZNF184_S71_L008_R1_001.B06_KB24_ZNF184_1… | RIP-Seq | 551.0 MB | link | |||
| — | KB25_293_CLIP_ZNF184_S72_L008_R1_001.C01_KB25_ZNF184_2… | RIP-Seq | 770.5 MB | link | |||
| — | KB25_293_CLIP_ZNF184_S72_L008_R1_001.D08fixed_KB25_ZNF… | RIP-Seq | 337.9 MB | link | |||
| — | KB3_293_CLIP_AIFM_2_IP_TCCGGAGA-ATAGAGGC_L008_R1.C01_K… | RIP-Seq | 343.0 MB | link | |||
| — | KB3_293_CLIP_AIFM_2_IP_TCCGGAGA-ATAGAGGC_L008_R1.D08fi… | RIP-Seq | 261.6 MB | link | |||
| — | KB5_293_CLIP_RNF219_1_input_ATTACTCG-AGGCGAAG_L008_R1.… | RIP-Seq | 656.7 MB | link | |||
| — | KB6_293_CLIP_RANGAP_3_IP_CGCTCATT-TATAGCCT_L008_R1.C01… | RIP-Seq | 219.5 MB | link | |||
| — | KB6_293_CLIP_RANGAP_3_IP_CGCTCATT-TATAGCCT_L008_R1.D08… | RIP-Seq | 230.1 MB | link | |||
| — | KB7_293_CLIP_RANGAP_4_IP_CGCTCATT-TAATCTTA_L008_R1.C01… | RIP-Seq | 229.1 MB | link | |||
| — | KB7_293_CLIP_RANGAP_4_IP_CGCTCATT-TAATCTTA_L008_R1.D08… | RIP-Seq | 175.2 MB | link | |||
| — | KB7_293_CLIP_RANGAP_4_IP_CGCTCATT-TAATCTTA_L008_R2.C01… | RIP-Seq | 229.1 MB | link | |||
| — | KB8_293_CLIP_RANGAP_2_input_ATTCAGAA-CCTATCCT_L008_R1.… | RIP-Seq | 704.3 MB | link | |||
| — | KB9_293_CLIP_NUMA_1_IP_GAATTCGT-GGCTCTGA_L008_R1.A01_K… | RIP-Seq | 139.5 MB | link | |||
| — | KB9_293_CLIP_NUMA_1_IP_GAATTCGT-GGCTCTGA_L008_R1.B06_K… | RIP-Seq | 219.8 MB | link | |||
| — | SRR4063774 | RIP-Seq | 704.3 MB | link | |||
| — | SRR4063789 | RIP-Seq | 433.9 MB | link |