GSE195494
GSE GEORibo-STAMP reveals translation status of RNAs bound by PABPN and PABPC
Relations
Summary
The poly(A) tail appended to the 3â 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- All reads were aligned to the human genome hg38 primary assembly using STAR.
- featureCounts version 2.0.2 was used to annotate reads, using the flag --minOverlap 20 and a custom GTF file derived from gencode v34
- EPKM values were determined as in (Brannan et al., 2021).
- 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.
- Control-STAMP PABPC IP Rep3 was not a successful immunoprecipitation and thus was excluded from analysis.
Supplementary Files (1)
Dataset Citations (1)
SRA Experiments (17) and Runs (17)
Total: 35574 MBSample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |
Sample attributes
Original files (1)
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 |