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GSE195495

GSE GEO
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Identification of different transcripts that favor the nuclear or cytoplasmic Poly(A) Binding Proteins (PABPs) through RNA immunoprecipitation

Organism: Homo sapiens
Platform: GPL20301
Samples: 9
Experiment Types:
Other
Submitted: Jan 26 2022
Last Updated: May 11 2022
Status: Public on Mar 30 2022
Contact: Amy,,Pasquinelli (University of California, San Diego)

Relations

BioProject: https://www.ncbi.nlm.nih.gov/bioproject/PRJNA800801

Summary

The poly(A) tail appended to the 3’ end of many eukaryotic transcripts plays a key role in the stability, nuclear transport, and translation of the RNA. These roles are largely mediated by the Poly(A) Binding Proteins (PABPs) that coat the poly(A) tails and interact with various proteins involved in the biogenesis and function of the RNA. While it is well established that the nuclear PABP (PABPN) binds newly synthesized poly(A) tails and is apparently replaced by the cytoplasmic PABP (PABPC) on transcripts exported to the cytoplasm, the distribution of transcripts for different genes or isoforms of the same gene on these distinct PABPs has not been investigated on a genome-wide scale. Here, we analyzed the identity, splicing status, poly(A) tail size, and translation status of RNAs co-immunoprecipitated with endogenous PABPN or PABPC in human cells. At steady state, many protein coding and non-coding RNAs exhibit strong bias for association with PABPN or PABPC. While PABPN-enriched transcripts more often were incompletely spliced and harbored longer poly(A) tails and PABPC-enriched RNAs tended to have longer half-lives and higher translation efficiency, there are curious outliers. Overall, our study reveals the landscape of RNAs bound by PABPN and PABPC, providing new details that support and advance the current understanding of the roles these proteins play in poly(A) tail synthesis, maintenance and function.

Overall Design

3 biological replicates were crosslinked and collected; a single input sample was divided so as to immunoprecipitate PABPN or PABPC from a single lysate sample. Antibodies used for immunoprecipitation were anti-PABPC, ab21060 Abcam; anti-PABPN, [EP3000Y] ab75855 Abcam.

Analysis (12 steps)

View Data Processing
Processing steps for GSE195495
  1. All reads were aligned to the human genome hg38 primary assembly using STAR.
  2. featureCounts version 2.0.2 was used to annotate reads, using the flag --minOverlap 20 and a custom GTF file derived from gencode v34
  3. Differential expression was calculated using DESeq2.
  4. Cut-offs used to determine significant differential expression were baseMean > 50 and padj <=0.01
  5. Genes which had substantial upstream intergenic reads (similar to DoGs) were removed from DE tables.
  6. Upstream intergenic regions were extracted using bedtools flank -l 2000 -r 0 -s.
  7. Intervening upstream genes that had any overlap with this region were removed with bedtools subtract.
  8. Reads that did not align to intronic or exonic regions were extracted from bam files with fgrep.
Showing first 8 steps.

Supplementary Files (2)

GSE195495_PABPCvInput_DE.csv.gz Download
GSE195495_PABPNvInput_DE.csv.gz Download
GEO Samples (9)

Dataset Citations (1)

Nuclear and cytoplasmic poly(A) binding proteins (PABPs) favor distinct transcripts and isoforms.
PMID 35438785 · 2022 · Nucleic acids research
Angela L Nicholson-Shaw, Eric R Kofman, Gene W Yeo, Amy E Pasquinelli

SRA Experiments (9) and Runs (9)

Total: 14916 MB
SRX13933701 SRP356890 RIP-Seq SINGLE
GSM5839370: HEK293T- Input RNA Rep1; Homo sapiens; RIP-Seq
Sample: SRS11777602
BioProject: PRJNA800801
BioSample: SAMN25284460
Platform: ILLUMINA
Instrument: Illumina HiSeq 4000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Total cell lysate
cell line: HEK293T
genotype: wildtype
treatment: formaldehyde crosslinked
immunoprecipitation: none
Original files (1)
HEK293T Total cell lysate
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771371 60576500 4603814000 1798.36 TotalInput_Rep1.fastq.gz, SRR17771371, SRR17771371.lite SRA
SRX13933702 SRP356890 RIP-Seq SINGLE
GSM5839371: HEK293T- Input RNA Rep2; Homo sapiens; RIP-Seq
Sample: SRS11777603
BioProject: PRJNA800801
BioSample: SAMN25284459
Platform: ILLUMINA
Instrument: Illumina HiSeq 4000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Total cell lysate
cell line: HEK293T
genotype: wildtype
treatment: formaldehyde crosslinked
immunoprecipitation: none
Original files (1)
HEK293T Total cell lysate
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771370 56793463 4316303188 1690.08 TotalInput_Rep2.fastq.gz, SRR17771370, SRR17771370.lite SRA
SRX13933703 SRP356890 RIP-Seq SINGLE
GSM5839372: HEK293T- Input RNA Rep3; Homo sapiens; RIP-Seq
Sample: SRS11777604
BioProject: PRJNA800801
BioSample: SAMN25284458
Platform: ILLUMINA
Instrument: Illumina HiSeq 4000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Total cell lysate
cell line: HEK293T
genotype: wildtype
treatment: formaldehyde crosslinked
immunoprecipitation: none
Original files (1)
HEK293T Total cell lysate
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771369 58036086 4410742536 1730.28 TotalInput_Rep3.fastq.gz, SRR17771369, SRR17771369.lite SRA
SRX13933704 SRP356890 RIP-Seq SINGLE
GSM5839373: HEK293T - PABPN RIP Rep1; Homo sapiens; RIP-Seq
Sample: SRS11777605
BioProject: PRJNA800801
BioSample: SAMN25284457
Platform: ILLUMINA
Instrument: Illumina HiSeq 4000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Total cell lysate
cell line: HEK293T
genotype: wildtype
treatment: formaldehyde crosslinked
immunoprecipitation: PABPN
Original files (1)
HEK293T Total cell lysate
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771368 51484132 3912794032 1525.25 RIP_PABPN_Rep1.fastq.gz, SRR17771368, SRR17771368.lite SRA
SRX13933705 SRP356890 RIP-Seq SINGLE
GSM5839374: HEK293T - PABPN RIP Rep2; Homo sapiens; RIP-Seq
Sample: SRS11777606
BioProject: PRJNA800801
BioSample: SAMN25284456
Platform: ILLUMINA
Instrument: Illumina HiSeq 4000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Total cell lysate
cell line: HEK293T
genotype: wildtype
treatment: formaldehyde crosslinked
immunoprecipitation: PABPN
Original files (1)
HEK293T Total cell lysate
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771367 59616577 4530859852 1770.71 RIP_PABPN_Rep2.fastq.gz, SRR17771367, SRR17771367.lite SRA
SRX13933706 SRP356890 RIP-Seq SINGLE
GSM5839375: HEK293T - PABPN RIP Rep3; Homo sapiens; RIP-Seq
Sample: SRS11777607
BioProject: PRJNA800801
BioSample: SAMN25284455
Platform: ILLUMINA
Instrument: Illumina HiSeq 4000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Total cell lysate
cell line: HEK293T
genotype: wildtype
treatment: formaldehyde crosslinked
immunoprecipitation: PABPN
Original files (1)
HEK293T Total cell lysate
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771366 55752851 4237216676 1658.61 RIP_PABPN_Rep3.fastq.gz, SRR17771366, SRR17771366.lite SRA
SRX13933707 SRP356890 RIP-Seq SINGLE
GSM5839376: HEK293T - PABPC RIP Rep1; Homo sapiens; RIP-Seq
Sample: SRS11777608
BioProject: PRJNA800801
BioSample: SAMN25284454
Platform: ILLUMINA
Instrument: Illumina HiSeq 4000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Total cell lysate
cell line: HEK293T
genotype: wildtype
treatment: formaldehyde crosslinked
immunoprecipitation: PABPC
Original files (1)
HEK293T Total cell lysate
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771365 58395305 4438043180 1905.57 RIP_PABPC_Rep1.fastq.gz, SRR17771365, SRR17771365.lite SRA
SRX13933708 SRP356890 RIP-Seq SINGLE
GSM5839377: HEK293T - PABPC RIP Rep2; Homo sapiens; RIP-Seq
Sample: SRS11777609
BioProject: PRJNA800801
BioSample: SAMN25284453
Platform: ILLUMINA
Instrument: Illumina HiSeq 4000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Total cell lysate
cell line: HEK293T
genotype: wildtype
treatment: formaldehyde crosslinked
immunoprecipitation: PABPC
Original files (1)
HEK293T Total cell lysate
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771364 44018523 3345407748 1318.0 RIP_PABPC_Rep2.fastq.gz, SRR17771364, SRR17771364.lite SRA
SRX13933709 SRP356890 RIP-Seq SINGLE
GSM5839378: HEK293T - PABPC RIP Rep3; Homo sapiens; RIP-Seq
Sample: SRS11777610
BioProject: PRJNA800801
BioSample: SAMN25284452
Platform: ILLUMINA
Instrument: Illumina HiSeq 4000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Total cell lysate
cell line: HEK293T
genotype: wildtype
treatment: formaldehyde crosslinked
immunoprecipitation: PABPC
Original files (1)
HEK293T Total cell lysate
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771363 50662493 3850349468 1519.4 RIP_PABPC_Rep3.fastq.gz, SRR17771363, SRR17771363.lite SRA

Linked Publications (2)

Data Files (9)

Accession File Name Stored Type Output Type Mapping Assembly Size Download
RIP_PABPC_Rep1.fastq.gz RIP-Seq 1.9 GB link
RIP_PABPC_Rep2.fastq.gz RIP-Seq 1.3 GB link
RIP_PABPC_Rep3.fastq.gz RIP-Seq 1.5 GB link
RIP_PABPN_Rep1.fastq.gz RIP-Seq 1.5 GB link
RIP_PABPN_Rep2.fastq.gz RIP-Seq 1.7 GB link
RIP_PABPN_Rep3.fastq.gz RIP-Seq 1.6 GB link
TotalInput_Rep1.fastq.gz RIP-Seq 1.8 GB link
TotalInput_Rep2.fastq.gz RIP-Seq 1.7 GB link
TotalInput_Rep3.fastq.gz RIP-Seq 1.7 GB link