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GSE195494

GSE GEO
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Ribo-STAMP reveals translation status of RNAs bound by PABPN and PABPC

Organism: Homo sapiens
Platform: GPL24676
Samples: 17
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/PRJNA800802

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, each replicate consists of a Control-STAMP sample and a RPS2-STAMP sample. These were crosslinked and collected; each 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 (5 steps)

View Data Processing
Processing steps for GSE195494
  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. EPKM values were determined as in (Brannan et al., 2021).
  4. Furthermore, for each gene, EPKM values obtained in the Control-STAMP cell line were subtracted from the RPS2-STAMP EPKM values, giving a normalized EPKM value for each gene which indicates the degree of translation of that transcript.
  5. Control-STAMP PABPC IP Rep3 was not a successful immunoprecipitation and thus was excluded from analysis.

Supplementary Files (1)

GSE195494_RiboSTAMP_norm_epkm.csv.gz Download
GEO Samples (17)

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 (17) and Runs (17)

Total: 35574 MB
SRX13933866 SRP356894 RIP-Seq PAIRED
GSM5839353: Control-STAMP Input RNA Rep1; Homo sapiens; RIP-Seq
Sample: SRS11777768
BioProject: PRJNA800802
BioSample: SAMN25284477
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Control-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced Control-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: none
Original files (1)
HEK293T Control-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771544 30230531 6106567262 1957.73 Input_CntrlSTAMP_Rep1_R1.fastq.gz, Input_CntrlSTAMP_Rep1_R2.fastq.gz,… SRA
SRX13933867 SRP356894 RIP-Seq PAIRED
GSM5839354: Control-STAMP Input RNA Rep2; Homo sapiens; RIP-Seq
Sample: SRS11777767
BioProject: PRJNA800802
BioSample: SAMN25284476
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Control-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced Control-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: none
Original files (1)
HEK293T Control-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771543 33830090 6833678180 2203.46 Input_CntrlSTAMP_Rep2_R1.fastq.gz, Input_CntrlSTAMP_Rep2_R2.fastq.gz,… SRA
SRX13933868 SRP356894 RIP-Seq PAIRED
GSM5839355: Control-STAMP Input RNA Rep3; Homo sapiens; RIP-Seq
Sample: SRS11777770
BioProject: PRJNA800802
BioSample: SAMN25284475
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Control-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced Control-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: none
Original files (1)
HEK293T Control-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771542 35404154 7151639108 2278.11 Input_CntrlSTAMP_Rep3_R1.fastq.gz, Input_CntrlSTAMP_Rep3_R2.fastq.gz,… SRA
SRX13933869 SRP356894 RIP-Seq PAIRED
GSM5839356: Control-STAMP IP PABPN RNA Rep1; Homo sapiens; RIP-Seq
Sample: SRS11777769
BioProject: PRJNA800802
BioSample: SAMN25284474
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Control-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced Control-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPN
Original files (1)
HEK293T Control-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771541 30001996 6060403192 1982.84 IP_PABPN_CntrlSTAMP_Rep1_R1.fastq.gz, IP_PABPN_CntrlSTAMP_Rep1_R2.fas… SRA
SRX13933870 SRP356894 RIP-Seq PAIRED
GSM5839357: Control-STAMP IP PABPN RNA Rep2; Homo sapiens; RIP-Seq
Sample: SRS11777771
BioProject: PRJNA800802
BioSample: SAMN25284473
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Control-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced Control-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPN
Original files (1)
HEK293T Control-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771540 32378271 6540410742 2101.74 IP_PABPN_CntrlSTAMP_Rep2_R1.fastq.gz, IP_PABPN_CntrlSTAMP_Rep2_R2.fas… SRA
SRX13933871 SRP356894 RIP-Seq PAIRED
GSM5839358: Control-STAMP IP PABPN RNA Rep3; Homo sapiens; RIP-Seq
Sample: SRS11777772
BioProject: PRJNA800802
BioSample: SAMN25284472
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Control-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced Control-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPN
Original files (1)
HEK293T Control-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771539 27977443 5651443486 1814.48 IP_PABPN_CntrlSTAMP_Rep3_R1.fastq.gz, IP_PABPN_CntrlSTAMP_Rep3_R2.fas… SRA
SRX13933872 SRP356894 RIP-Seq PAIRED
GSM5839359: Control-STAMP IP PABPC RNA Rep1; Homo sapiens; RIP-Seq
Sample: SRS11777773
BioProject: PRJNA800802
BioSample: SAMN25284471
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Control-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced Control-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPC
Original files (1)
HEK293T Control-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771538 29856052 6030922504 1939.89 IP_PABPC_CntrlSTAMP_Rep1_R1.fastq.gz, IP_PABPC_CntrlSTAMP_Rep1_R2.fas… SRA
SRX13933873 SRP356894 RIP-Seq PAIRED
GSM5839360: Control-STAMP IP PABPC RNA Rep2; Homo sapiens; RIP-Seq
Sample: SRS11777774
BioProject: PRJNA800802
BioSample: SAMN25284470
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T Control-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced Control-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPC
Original files (1)
HEK293T Control-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771537 33262542 6719033484 2137.79 IP_PABPC_CntrlSTAMP_Rep2_R1.fastq.gz, IP_PABPC_CntrlSTAMP_Rep2_R2.fas… SRA
SRX13933874 SRP356894 RIP-Seq PAIRED
GSM5839361: RPS2-STAMP Input RNA Rep1; Homo sapiens; RIP-Seq
Sample: SRS11777775
BioProject: PRJNA800802
BioSample: SAMN25284469
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T RPS2-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced RPS2-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: none
Original files (1)
HEK293T RPS2-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771536 30132567 6086778534 1963.82 Input_RPS2STAMP_Rep1_R1.fastq.gz, Input_RPS2STAMP_Rep1_R2.fastq.gz, S… SRA
SRX13933875 SRP356894 RIP-Seq PAIRED
GSM5839362: RPS2-STAMP Input RNA Rep2; Homo sapiens; RIP-Seq
Sample: SRS11777776
BioProject: PRJNA800802
BioSample: SAMN25284468
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T RPS2-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced RPS2-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: none
Original files (1)
HEK293T RPS2-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771534 31138645 6290006290 2005.08 Input_RPS2STAMP_Rep2_R1.fastq.gz, Input_RPS2STAMP_Rep2_R2.fastq.gz, S… SRA
SRX13933876 SRP356894 RIP-Seq PAIRED
GSM5839363: RPS2-STAMP Input RNA Rep3; Homo sapiens; RIP-Seq
Sample: SRS11777777
BioProject: PRJNA800802
BioSample: SAMN25284467
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T RPS2-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced RPS2-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: none
Original files (1)
HEK293T RPS2-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771533 34238614 6916200028 2226.48 Input_RPS2STAMP_Rep3_R1.fastq.gz, Input_RPS2STAMP_Rep3_R2.fastq.gz, S… SRA
SRX13933877 SRP356894 RIP-Seq PAIRED
GSM5839364: RPS2-STAMP IP PABPN RNA Rep1; Homo sapiens; RIP-Seq
Sample: SRS11777778
BioProject: PRJNA800802
BioSample: SAMN25284466
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T RPS2-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced RPS2-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPN
Original files (1)
HEK293T RPS2-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771532 35073782 7084903964 2294.13 IP_PABPN_RPS2STAMP_Rep1_R1.fastq.gz, IP_PABPN_RPS2STAMP_Rep1_R2.fastq… SRA
SRX13933878 SRP356894 RIP-Seq PAIRED
GSM5839365: RPS2-STAMP IP PABPN RNA Rep2; Homo sapiens; RIP-Seq
Sample: SRS11777779
BioProject: PRJNA800802
BioSample: SAMN25284465
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T RPS2-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced RPS2-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPN
Original files (1)
HEK293T RPS2-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771535 31351160 6332934320 2039.4 IP_PABPN_RPS2STAMP_Rep2_R1.fastq.gz, IP_PABPN_RPS2STAMP_Rep2_R2.fastq… SRA
SRX13933879 SRP356894 RIP-Seq PAIRED
GSM5839366: RPS2-STAMP IP PABPN RNA Rep3; Homo sapiens; RIP-Seq
Sample: SRS11777780
BioProject: PRJNA800802
BioSample: SAMN25284464
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T RPS2-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced RPS2-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPN
Original files (1)
HEK293T RPS2-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771531 42395522 8563895444 2776.14 IP_PABPN_RPS2STAMP_Rep3_R1.fastq.gz, IP_PABPN_RPS2STAMP_Rep3_R2.fastq… SRA
SRX13933880 SRP356894 RIP-Seq PAIRED
GSM5839367: RPS2-STAMP IP PABPC RNA Rep1; Homo sapiens; RIP-Seq
Sample: SRS11777781
BioProject: PRJNA800802
BioSample: SAMN25284463
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T RPS2-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced RPS2-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPC
Original files (1)
HEK293T RPS2-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771530 26264063 5305340726 1684.16 IP_PABPC_RPS2STAMP_Rep1_R1.fastq.gz, IP_PABPC_RPS2STAMP_Rep1_R2.fastq… SRA
SRX13933881 SRP356894 RIP-Seq PAIRED
GSM5839368: RPS2-STAMP IP PABPC RNA Rep2; Homo sapiens; RIP-Seq
Sample: SRS11777782
BioProject: PRJNA800802
BioSample: SAMN25284462
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T RPS2-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced RPS2-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPC
Original files (1)
HEK293T RPS2-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771529 32502244 6565453288 2083.41 IP_PABPC_RPS2STAMP_Rep2_R1.fastq.gz, IP_PABPC_RPS2STAMP_Rep2_R2.fastq… SRA
SRX13933882 SRP356894 RIP-Seq PAIRED
GSM5839369: RPS2-STAMP IP PABPC RNA Rep3; Homo sapiens; RIP-Seq
Sample: SRS11777783
BioProject: PRJNA800802
BioSample: SAMN25284461
Platform: ILLUMINA
Instrument: Illumina NovaSeq 6000
Organism: Homo sapiens
Sample attributes
source_name: HEK293T RPS2-APOBEC1 Cell line
cell line: HEK293T
stamp condition: Induced RPS2-APOBEC1
treatment: formaldehyde crosslinked
immunoprecipitation: PABPC
Original files (1)
HEK293T RPS2-APOBEC1 Cell line
Runs (1)
Run Spots Bases Size (MB) Files Link
SRR17771528 32375157 6539781714 2085.45 IP_PABPC_RPS2STAMP_Rep3_R1.fastq.gz, IP_PABPC_RPS2STAMP_Rep3_R2.fastq… SRA

Linked Publications (2)

Data Files (17)

Accession File Name Stored Type Output Type Mapping Assembly Size Download
Input_CntrlSTAMP_Rep1_R1.fastq.gz RIP-Seq 1.9 GB link
Input_CntrlSTAMP_Rep2_R1.fastq.gz RIP-Seq 2.2 GB link
Input_CntrlSTAMP_Rep3_R1.fastq.gz RIP-Seq 2.2 GB link
Input_RPS2STAMP_Rep1_R1.fastq.gz RIP-Seq 1.9 GB link
Input_RPS2STAMP_Rep2_R1.fastq.gz RIP-Seq 2.0 GB link
Input_RPS2STAMP_Rep3_R1.fastq.gz RIP-Seq 2.2 GB link
IP_PABPC_CntrlSTAMP_Rep1_R1.fastq.gz RIP-Seq 1.9 GB link
IP_PABPC_CntrlSTAMP_Rep2_R1.fastq.gz RIP-Seq 2.1 GB link
IP_PABPC_RPS2STAMP_Rep1_R1.fastq.gz RIP-Seq 1.6 GB link
IP_PABPC_RPS2STAMP_Rep2_R1.fastq.gz RIP-Seq 2.0 GB link
IP_PABPC_RPS2STAMP_Rep3_R1.fastq.gz RIP-Seq 2.0 GB link
IP_PABPN_CntrlSTAMP_Rep1_R1.fastq.gz RIP-Seq 1.9 GB link
IP_PABPN_CntrlSTAMP_Rep2_R1.fastq.gz RIP-Seq 2.1 GB link
IP_PABPN_CntrlSTAMP_Rep3_R1.fastq.gz RIP-Seq 1.8 GB link
IP_PABPN_RPS2STAMP_Rep1_R1.fastq.gz RIP-Seq 2.2 GB link
IP_PABPN_RPS2STAMP_Rep2_R1.fastq.gz RIP-Seq 2.0 GB link
IP_PABPN_RPS2STAMP_Rep3_R1.fastq.gz RIP-Seq 2.7 GB link