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Large-scale tethered function assays identify factors that regulate mRNA stability and translation.

En-Ching Luo, Jason L Nathanson, Frederick E Tan, Joshua L Schwartz, Jonathan C Schmok, Archana Shankar, Sebastian Markmiller, Brian A Yee, Shashank Sathe, Gabriel A Pratt, Duy B Scaletta, Yuanchi Ha, David E Hill, Stefan Aigner, Gene W Yeo

Nature structural & molecular biology · 2020 · Vol. 27 (10) · pp. 989-1000

Abstract

The molecular functions of the majority of RNA-binding proteins (RBPs) remain unclear, highlighting a major bottleneck to a full understanding of gene expression regulation. Here, we develop a plasmid resource of 690 human RBPs that we subject to luciferase-based 3'-untranslated-region tethered function assays to pinpoint RBPs that regulate RNA stability or translation. Enhanced UV-cross-linking and immunoprecipitation of these RBPs identifies thousands of endogenous mRNA targets that respond to changes in RBP level, recapitulating effects observed in tethered function assays. Among these RBPs, the ubiquitin-associated protein 2-like (UBAP2L) protein interacts with RNA via its RGG domain and cross-links to mRNA and rRNA. Fusion of UBAP2L to RNA-targeting CRISPR-Cas9 demonstrates programmable translational enhancement. Polysome profiling indicates that UBAP2L promotes translation of target mRNAs, particularly global regulators of translation. Our tethering survey allows rapid assignment of the molecular activity of proteins, such as UBAP2L, to specific steps of mRNA metabolism.

Linked datasets 4

GSE117290 GSE · via ncbi_elink
GEO ↗

Large-scale tethered function assays identify factors that regulate mRNA stability and translation (eCLIP)

Homo sapiens

113 data files
FileTypeSize
EC_AIMP1_R1_IN_S41_L004_R1_001.fastq.gz OTHER 520.5 MB
EC_AIMP1_R1_IP_S43_L004_R1_001.fastq.gz OTHER 624.5 MB
EC_AIMP1_R2_IN_S42_L004_R1_001.fastq.gz OTHER 731.0 MB
EC_AIMP1_R2_IP_S44_L004_R1_001.fastq.gz OTHER 615.3 MB
EC_BOLL_R1_IN_S49_L005_R1_001.fastq.gz OTHER 716.2 MB
EC_BOLL_R1_IP_S51_L005_R1_001.fastq.gz OTHER 646.9 MB
EC_BOLL_R2_IN_S50_L005_R1_001.fastq.gz OTHER 780.6 MB
EC_BOLL_R2_IP_S52_L005_R1_001.fastq.gz OTHER 813.4 MB
EC_CLK3_R1_IN_S29_L003_R1_001.fastq.gz OTHER 525.5 MB
EC_CLK3_R1_IP_S31_L003_R1_001.fastq.gz OTHER 563.0 MB
EC_CLK3_R2_IN_S30_L003_R1_001.fastq.gz OTHER 436.9 MB
EC_CLK3_R2_IP_S32_L003_R1_001.fastq.gz OTHER 861.6 MB
EC_CNOT7_R1_IN_S53_L005_R1_001.fastq.gz OTHER 676.6 MB
EC_CNOT7_R1_IP_S55_L005_R1_001.fastq.gz OTHER 542.1 MB
EC_CNOT7_R2_IN_S54_L005_R1_001.fastq.gz OTHER 661.8 MB
EC_CNOT7_R2_IP_S56_L005_R1_001.fastq.gz OTHER 624.3 MB
EC_DDX6_R1-IN_S5_L001_R1_001.fastq.gz OTHER 870.3 MB
EC_DDx6_R1-IP_S7_L001_R1_001.fastq.gz OTHER 990.0 MB
EC_DDx6_R2-IN_S6_L001_R1_001.fastq.gz OTHER 809.9 MB
EC_DDX6_R2-IP_S8_L001_R1_001.fastq.gz OTHER 1.1 GB
EC_IFIT2_R1-IN_S9_L002_R1_001.fastq.gz OTHER 545.5 MB
EC_IFIT2_R1-IP_S13_L002_R1_001.fastq.gz OTHER 611.7 MB
EC_IFIT2_R2-IN_S10_L002_R1_001.fastq.gz OTHER 616.1 MB
EC_IFIT2_R2-IP_S14_L002_R1_001.fastq.gz OTHER 681.2 MB
EC_MEX3C_R1-IN_S21_L003_R1_001.fastq.gz OTHER 573.1 MB
EC_MEX3C_R1-IP_S23_L003_R1_001.fastq.gz OTHER 1.1 GB
EC_MEX3C_R2-IN_S22_L003_R1_001.fastq.gz OTHER 599.9 MB
EC_MEX3C_R2-IP_S24_L003_R1_001.fastq.gz OTHER 933.6 MB
EC_MTDH_R1_IN_S33_L004_R1_001.fastq.gz OTHER 782.9 MB
EC_MTDH_R1_IP_S35_L004_R1_001.fastq.gz OTHER 848.6 MB
EC_MTDH_R2_IN_S34_L004_R1_001.fastq.gz OTHER 853.8 MB
EC_MTDH_R2_IP_S36_L004_R1_001.fastq.gz OTHER 625.0 MB
EC_NANOS3_R1-IN_S17_L002_R1_001.fastq.gz OTHER 566.0 MB
EC_NANOS3_R1-IP_S19_L002_R1_001.fastq.gz OTHER 746.0 MB
EC_NANOS3_R2-IN_S18_L002_R1_001.fastq.gz OTHER 576.9 MB
EC_NANOS3_R2-IP_S20_L002_R1_001.fastq.gz OTHER 598.6 MB
EC_PARN_R1-IN_S25_L003_R1_001.fastq.gz OTHER 617.7 MB
EC_PARN_R1-IP_S27_L003_R1_001.fastq.gz OTHER 551.6 MB
EC_PARN_R2-IN_S26_L003_R1_001.fastq.gz OTHER 549.8 MB
EC_PARN_R2-IP_S28_L003_R1_001.fastq.gz OTHER 1.2 GB
EC_SNRPA_R1_IN_S45_L005_R1_001.fastq.gz OTHER 755.7 MB
EC_SNRPA_R1_IP_S47_L005_R1_001.fastq.gz OTHER 1.1 GB
EC_SNRPA_R2_IN_S46_L005_R1_001.fastq.gz OTHER 642.7 MB
EC_SNRPA_R2_IP_S48_L005_R1_001.fastq.gz OTHER 618.8 MB
EC_TBO2_R1-IN_S11_L002_R1_001.fastq.gz OTHER 564.3 MB
EC_TBO2_R1-IP_S15_L002_R1_001.fastq.gz OTHER 1.5 GB
EC_TBO2_R2-IN_S12_L002_R1_001.fastq.gz OTHER 428.8 MB
EC_TBO2_R2-IP_S16_L002_R1_001.fastq.gz OTHER 559.1 MB
EC_TOB1_R1_IN_S37_L004_R1_001.fastq.gz OTHER 691.3 MB
EC_TOB1_R1_IP_S39_L004_R1_001.fastq.gz OTHER 881.0 MB
EC_TOB1_R2_IN_S38_L004_R1_001.fastq.gz OTHER 720.5 MB
EC_TOB1_R2_IP_S40_L004_R1_001.fastq.gz OTHER 778.4 MB
EC_UBAP2L_R1_IN_S7_L007_R1_001.fastq.gz OTHER 755.7 MB
EC_UBAP2L_R1_IP_S8_L007_R1_001.fastq.gz OTHER 566.8 MB
EC_UBAP2L_R2_IN_S9_L007_R1_001.fastq.gz OTHER 1.2 GB
EC_UBAP2L_R2_IP_S10_L007_R1_001.fastq.gz OTHER 885.1 MB
SRR11683564_GSM4512262 OTHER 676.0 MB
SRR11683565_GSM4512263 OTHER 542.0 MB
SRR11683566_GSM4512264 OTHER 661.0 MB
SRR11683567_GSM4512265 OTHER 624.0 MB
SRR11683568_GSM4512266 OTHER 520.0 MB
SRR11683569_GSM4512267 OTHER 624.0 MB
SRR11683570_GSM4512268 OTHER 730.0 MB
SRR11683571_GSM4512269 OTHER 615.0 MB
SRR11683572_GSM4512270 OTHER 782.0 MB
SRR11683573_GSM4512271 OTHER 848.0 MB
SRR11683574_GSM4512272 OTHER 853.0 MB
SRR11683575_GSM4512273 OTHER 625.0 MB
SRR11683576_GSM4512274 OTHER 755.0 MB
SRR11683577_GSM4512275 OTHER 566.0 MB
SRR11683578_GSM4512276 OTHER 1.2 GB
SRR11683579_GSM4512277 OTHER 885.0 MB
SRR11683580_GSM4512278 OTHER 525.0 MB
SRR11683581_GSM4512279 OTHER 562.0 MB
SRR11683582_GSM4512280 OTHER 436.0 MB
SRR11683583_GSM4512281 OTHER 861.0 MB
SRR11683584_GSM4512282 OTHER 870.0 MB
SRR11683585_GSM4512283 OTHER 989.0 MB
SRR11683586_GSM4512284 OTHER 809.0 MB
SRR11683587_GSM4512285 OTHER 1.1 GB
SRR11683588_GSM4512286 OTHER 573.0 MB
SRR11683589_GSM4512287 OTHER 1.1 GB
SRR11683590_GSM4512288 OTHER 599.0 MB
SRR11683591_GSM4512289 OTHER 933.0 MB
SRR11683592_GSM4512290 OTHER 617.0 MB
SRR11683593_GSM4512291 OTHER 551.0 MB
SRR11683594_GSM4512292 OTHER 549.0 MB
SRR11683595_GSM4512293 OTHER 1.2 GB
SRR11683596_GSM4512294 OTHER 755.0 MB
SRR11683597_GSM4512295 OTHER 1.1 GB
SRR11683598_GSM4512296 OTHER 642.0 MB
SRR11683599_GSM4512297 OTHER 618.0 MB
SRR11683600_GSM4512298 OTHER 564.0 MB
SRR11683601_GSM4512299 OTHER 1.5 GB
SRR11683602_GSM4512300 OTHER 428.0 MB
SRR11683603_GSM4512301 OTHER 559.0 MB
SRR11683604_GSM4512302 OTHER 716.0 MB
SRR11683605_GSM4512303 OTHER 646.0 MB
SRR11683606_GSM4512304 OTHER 780.0 MB
SRR11683607_GSM4512305 OTHER 813.0 MB
SRR11683608_GSM4512306 OTHER 545.0 MB
SRR11683609_GSM4512307 OTHER 611.0 MB
SRR11683610_GSM4512308 OTHER 616.0 MB
SRR11683611_GSM4512309 OTHER 681.0 MB
SRR11683611.lite OTHER 681.2 MB
SRR11683612_GSM4512310 OTHER 565.0 MB
SRR11683613_GSM4512311 OTHER 745.0 MB
SRR11683614_GSM4512312 OTHER 576.0 MB
SRR11683615_GSM4512313 OTHER 598.0 MB
SRR11683616_GSM4512314 OTHER 691.0 MB
SRR11683617_GSM4512315 OTHER 881.0 MB
SRR11683618_GSM4512316 OTHER 720.0 MB
SRR11683619_GSM4512317 OTHER 778.0 MB
GSE117291 GSE · via ncbi_elink
GEO ↗

Large-scale tethered function assays identify factors that regulate mRNA stability and translation (polysome profiling)

Homo sapiens

24 data files
FileTypeSize
EC_Input_293T_1_S1_L001_R1_001.fastq.gz RNA-Seq 1.6 GB
EC_Input_293T_2_S2_L001_R1_001.fastq.gz RNA-Seq 1.7 GB
EC_Input_KO1_1_S3_L001_R1_001.fastq.gz RNA-Seq 873.8 MB
EC_Input_KO1_2_S4_L001_R1_001.fastq.gz RNA-Seq 527.9 MB
EC_Input_KO9_1_S5_L001_R1_001.fastq.gz RNA-Seq 1016.5 MB
EC_Input_KO9_2_S6_L001_R1_001.fastq.gz RNA-Seq 723.3 MB
EC_Poly_293T_1_S7_L001_R1_001.fastq.gz RNA-Seq 893.3 MB
EC_Poly_293T_2_S8_L001_R1_001.fastq.gz RNA-Seq 534.6 MB
EC_Poly_KO1_1_S9_L001_R1_001.fastq.gz RNA-Seq 665.6 MB
EC_Poly_KO1_2_S10_L001_R1_001.fastq.gz RNA-Seq 624.7 MB
EC_Poly_KO9_1_S11_L001_R1_001.fastq.gz RNA-Seq 222.8 MB
EC_Poly_KO9_2_S12_L001_R1_001.fastq.gz RNA-Seq 395.0 MB
SRR7547726_GSM3290512 RNA-Seq 1.6 GB
SRR7547727_GSM3290513 RNA-Seq 1.7 GB
SRR7547728_GSM3290514 RNA-Seq 873.0 MB
SRR7547729_GSM3290515 RNA-Seq 527.0 MB
SRR7547730_GSM3290516 RNA-Seq 1016.0 MB
SRR7547731_GSM3290517 RNA-Seq 723.0 MB
SRR7547732_GSM3290518 RNA-Seq 893.0 MB
SRR7547733_GSM3290519 RNA-Seq 534.0 MB
SRR7547734_GSM3290520 RNA-Seq 665.0 MB
SRR7547735_GSM3290521 RNA-Seq 624.0 MB
SRR7547736_GSM3290522 RNA-Seq 222.0 MB
SRR7547737_GSM3290523 RNA-Seq 394.0 MB
GSE117294 GSE · via ncbi_elink
GEO ↗

Large-scale tethered function assays identify factors that regulate mRNA stability and translation

Homo sapiens

GSE117293 GSE · via ncbi_elink
GEO ↗

Large-scale tethered function assays identify factors that regulate mRNA stability and translation (HEK293T_RNA-seq)

Homo sapiens

100 data files
FileTypeSize
EC-KD-s1-CLK3-1-1_S7_L001_R1_001.fastq.gz RNA-Seq 647.4 MB
EC-KD-s1-CLK3-1-2_S8_L001_R1_001.fastq.gz RNA-Seq 597.8 MB
EC-KD-s1-CNOT7-1-1_S3_L001_R1_001.fastq.gz RNA-Seq 532.8 MB
EC-KD-s1-CNOT7-1-2_S4_L001_R1_001.fastq.gz RNA-Seq 603.4 MB
EC-KD-s1-CNOT7-2-1_S5_L001_R1_001.fastq.gz RNA-Seq 480.6 MB
EC-KD-s1-CNOT7-2-2_S6_L001_R1_001.fastq.gz RNA-Seq 536.8 MB
EC-KD-s1-ctrl-2_S1_L001_R1_001.fastq.gz RNA-Seq 515.5 MB
EC-KD-s1-ctrl-3_S2_L001_R1_001.fastq.gz RNA-Seq 524.1 MB
EC-KD-s1-UBAP2L-1-1_S9_L001_R1_001.fastq.gz RNA-Seq 521.0 MB
EC-KD-s1-UBAP2L-1-2_S10_L001_R1_001.fastq.gz RNA-Seq 499.4 MB
EC-KD-s1-UBAP2L-2-1_S11_L001_R1_001.fastq.gz RNA-Seq 496.5 MB
EC-KD-s1-UBAP2L-2-2_S12_L001_R1_001.fastq.gz RNA-Seq 477.8 MB
EC-KD-s2-AIMP1-1-1_S13_L002_R1_001.fastq.gz RNA-Seq 882.9 MB
EC-KD-s2-AIMP1-1-2_S14_L002_R1_001.fastq.gz RNA-Seq 534.1 MB
EC-KD-s2-AIMP1-2-1_S15_L002_R1_001.fastq.gz RNA-Seq 428.5 MB
EC-KD-s2-AIMP1-2-2_S16_L002_R1_001.fastq.gz RNA-Seq 562.2 MB
EC-KD-s2-ctrl-1-1_S25_L003_R1_001.fastq.gz RNA-Seq 512.6 MB
EC-KD-s2-ctrl-1-2_S26_L003_R1_001.fastq.gz RNA-Seq 509.2 MB
EC-KD-s2-DDX6-1-1_S17_L002_R1_001.fastq.gz RNA-Seq 481.4 MB
EC-KD-s2-DDX6-1-2_S18_L002_R1_001.fastq.gz RNA-Seq 551.1 MB
EC-KD-s2-DDX6-2-1_S19_L002_R1_001.fastq.gz RNA-Seq 590.2 MB
EC-KD-s2-DDX6-2-2_S20_L002_R1_001.fastq.gz RNA-Seq 438.8 MB
EC-KD-s2-MEX3C-1-1_S21_L002_R1_001.fastq.gz RNA-Seq 383.7 MB
EC-KD-s2-MEX3C-1-2_S22_L002_R1_001.fastq.gz RNA-Seq 647.6 MB
EC-KD-s2-MEX3C-2-1_S23_L002_R1_001.fastq.gz RNA-Seq 340.8 MB
EC-KD-s2-MEX3C-2-2_S24_L002_R1_001.fastq.gz RNA-Seq 569.3 MB
EC-KD-s3-ctrl-1-1_S39_L004_R1_001.fastq.gz RNA-Seq 678.9 MB
EC-KD-s3-ctrl-1-2_S40_L004_R1_001.fastq.gz RNA-Seq 563.8 MB
EC-KD-s3-MTDH-1-1_S27_L003_R1_001.fastq.gz RNA-Seq 390.9 MB
EC-KD-s3-MTDH-1-2_S28_L003_R1_001.fastq.gz RNA-Seq 425.8 MB
EC-KD-s3-MTDH-2-1_S29_L003_R1_001.fastq.gz RNA-Seq 664.9 MB
EC-KD-s3-MTDH-2-2_S30_L003_R1_001.fastq.gz RNA-Seq 619.9 MB
EC-KD-s3-PARN-1-1_S31_L003_R1_001.fastq.gz RNA-Seq 322.0 MB
EC-KD-s3-PARN-1-2_S32_L003_R1_001.fastq.gz RNA-Seq 479.3 MB
EC-KD-s3-PARN-2-1_S33_L003_R1_001.fastq.gz RNA-Seq 564.2 MB
EC-KD-s3-PARN-2-2_S34_L003_R1_001.fastq.gz RNA-Seq 660.1 MB
EC-OE-BOLL-1_S49_L005_R1_001.fastq.gz RNA-Seq 622.9 MB
EC-OE-BOLL-2_S50_L005_R1_001.fastq.gz RNA-Seq 545.8 MB
EC-OE-FLAG-1_S57_L005_R1_001.fastq.gz RNA-Seq 713.2 MB
EC-OE-FLAG-2_S58_L005_R1_001.fastq.gz RNA-Seq 678.1 MB
EC-OE-IFIT2-1_S43_L004_R1_001.fastq.gz RNA-Seq 533.2 MB
EC-OE-IFIT2-2_S44_L004_R1_001.fastq.gz RNA-Seq 560.3 MB
EC-OE-NANOS3-1_S41_L004_R1_001.fastq.gz RNA-Seq 617.8 MB
EC-OE-NANOS3-2_S42_L004_R1_001.fastq.gz RNA-Seq 532.1 MB
EC-OE-SNRPA-1_S55_L005_R1_001.fastq.gz RNA-Seq 688.4 MB
EC-OE-SNRPA-2_S56_L005_R1_001.fastq.gz RNA-Seq 676.4 MB
EC-OE-TOB1-1_S51_L005_R1_001.fastq.gz RNA-Seq 771.7 MB
EC-OE-TOB1-2_S52_L005_R1_001.fastq.gz RNA-Seq 568.2 MB
EC-OE-TOB2-1_S53_L005_R1_001.fastq.gz RNA-Seq 601.9 MB
EC-OE-TOB2-2_S54_L005_R1_001.fastq.gz RNA-Seq 641.4 MB
SRR7547754_GSM3290532 RNA-Seq 647.0 MB
SRR7547755_GSM3290533 RNA-Seq 597.0 MB
SRR7547756_GSM3290534 RNA-Seq 532.0 MB
SRR7547757_GSM3290535 RNA-Seq 603.0 MB
SRR7547758_GSM3290536 RNA-Seq 480.0 MB
SRR7547759_GSM3290537 RNA-Seq 536.0 MB
SRR7547760_GSM3290538 RNA-Seq 515.0 MB
SRR7547761_GSM3290539 RNA-Seq 524.0 MB
SRR7547762_GSM3290540 RNA-Seq 521.0 MB
SRR7547763_GSM3290541 RNA-Seq 499.0 MB
SRR7547764_GSM3290542 RNA-Seq 496.0 MB
SRR7547765_GSM3290543 RNA-Seq 477.0 MB
SRR7547766_GSM3290544 RNA-Seq 882.0 MB
SRR7547767_GSM3290545 RNA-Seq 534.0 MB
SRR7547768_GSM3290546 RNA-Seq 428.0 MB
SRR7547769_GSM3290547 RNA-Seq 562.0 MB
SRR7547770_GSM3290548 RNA-Seq 512.0 MB
SRR7547771_GSM3290549 RNA-Seq 509.0 MB
SRR7547772_GSM3290550 RNA-Seq 481.0 MB
SRR7547773_GSM3290551 RNA-Seq 551.0 MB
SRR7547774_GSM3290552 RNA-Seq 590.0 MB
SRR7547775_GSM3290553 RNA-Seq 438.0 MB
SRR7547776_GSM3290554 RNA-Seq 383.0 MB
SRR7547777_GSM3290555 RNA-Seq 647.0 MB
SRR7547778_GSM3290556 RNA-Seq 340.0 MB
SRR7547779_GSM3290557 RNA-Seq 569.0 MB
SRR7547780_GSM3290558 RNA-Seq 678.0 MB
SRR7547781_GSM3290559 RNA-Seq 563.0 MB
SRR7547782_GSM3290560 RNA-Seq 390.0 MB
SRR7547783_GSM3290561 RNA-Seq 425.0 MB
SRR7547784_GSM3290562 RNA-Seq 664.0 MB
SRR7547785_GSM3290563 RNA-Seq 619.0 MB
SRR7547786_GSM3290564 RNA-Seq 322.0 MB
SRR7547787_GSM3290565 RNA-Seq 479.0 MB
SRR7547788_GSM3290566 RNA-Seq 564.0 MB
SRR7547789_GSM3290567 RNA-Seq 660.0 MB
SRR7547790_GSM3290568 RNA-Seq 622.0 MB
SRR7547791_GSM3290569 RNA-Seq 545.0 MB
SRR7547792_GSM3290570 RNA-Seq 713.0 MB
SRR7547793_GSM3290571 RNA-Seq 678.0 MB
SRR7547794_GSM3290572 RNA-Seq 533.0 MB
SRR7547795_GSM3290573 RNA-Seq 560.0 MB
SRR7547796_GSM3290574 RNA-Seq 617.0 MB
SRR7547797_GSM3290575 RNA-Seq 532.0 MB
SRR7547798_GSM3290576 RNA-Seq 688.0 MB
SRR7547799_GSM3290577 RNA-Seq 676.0 MB
SRR7547800_GSM3290578 RNA-Seq 771.0 MB
SRR7547801_GSM3290579 RNA-Seq 568.0 MB
SRR7547802_GSM3290580 RNA-Seq 601.0 MB
SRR7547803_GSM3290581 RNA-Seq 641.0 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.

Accession Source Title
ENCSR456FVU ENCSR · ENCODE No title in the index
GSE117294 GSE · GEO GEO ↗ Large-scale tethered function assays identify factors that regulate mRNA stability and translation

Analysis pipelines 4

Pipeline Assay Steps Dataset
GSE117290 eCLIP Data Processing geo_data_processing eCLIP 7 GSE117290
GSE117291 RNA-seq Data Processing geo_data_processing RNA-seq 5 GSE117291
GSE117293 RNA-seq Data Processing geo_data_processing RNA-seq 5 GSE117293
GSE117294 Data Processing geo_data_processing — 5 GSE117294
Funding 4 grants
Grant Funder
R01 HG004659 NHGRI NIH HHS (United States)
R01 NS103172 NINDS NIH HHS (United States)
U41 HG009889 NHGRI NIH HHS (United States)
U19 MH107367 NIMH NIH HHS (United States)