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| Status |
Public on Jul 15, 2013 |
| Title |
Global signatures of protein binding on structural RNAs in Saccharomyces cerevisiae |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Protein binding is essential to the transport, decay and regulation of almost all RNA molecules. However, the structural preference of protein binding on RNAs and their cellelar functions and dynamics upon changing environmental condictions are poorly understood. Here, we integrated various high-throughput data and introduced a computational framework to describe the global interactions between RNA binding proteins (RBPs) and structured RNAs in yeast at single-nucleotide resolution. We found that on average, in terms of percent total lengths, ~15% of mRNA untranslated regions (UTRs), ~37% of canonical ncRNAs and ~11% of long ncRNA (lncRNAs) are bound by proteins. The RBP binding sites, in general, tend to occur at single-stranded loops, with evolutionarily conserved signatures, and often facilitate a specific RNA structure conformation in vivo. We found that four nucleotide modifications of tRNA are significantly associated with RBP binding. We also identified various structural motifs bound by RBPs in the UTRs of mRNAs, associated with localization, degradation and stress responces. Moreover, we identified >200 novel lncRNAs bound by RBPs, and about half of them contain conserved secondary structures. We present the first ensemble pattern of RBP binding sites in the structured noncoding regions of a eukaryotic genome, emphasizing their structural context and cellular functions.
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| Overall design |
Duplicate gPAR-CLIP libraries were sequenced from yeast strains for each of three conditions: log-phase growth, growth after 2 hour glucose starvation, and growth after 2 hour nitrogen starvation. polyA RNAs were isolated for all conditions.
Total RNA were isolated from log phase growth conditions. Sucrose gradient fractionation was performed: some RNAs were isolated from the "light" fraction (lighter than 40S ribosome) and some from the "heavy" fraction. gPAR-CLIP libraries were used to determine regions of RNA bound by proteins.
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| Contributor(s) |
Freeberg MA, Han T, Yang Y, Lu Z |
| Citation(s) |
24369346 |
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| Submission date |
Jul 15, 2013 |
| Last update date |
May 15, 2019 |
| Contact name |
Mallory Ann Freeberg |
| E-mail(s) |
mfreebe2@jhu.edu
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| Organization name |
Johns Hopkins University
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| Department |
Department of Biology
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| Street address |
3400 N Charles St
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| City |
Baltimore |
| State/province |
MD |
| ZIP/Postal code |
21211 |
| Country |
USA |
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| Platforms (1) |
| GPL13821 |
Illumina HiSeq 2000 (Saccharomyces cerevisiae) |
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| Samples (10)
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| GSM1186342 |
SD, total RNA, light fraction, gPAR-CLIP, Replicate 1 |
| GSM1186343 |
SD, total RNA, light fraction, gPAR-CLIP, Replicate 2 |
| GSM1186344 |
SD, total RNA, heavy fraction, gPAR-CLIP, Replicate 1 |
| GSM1186345 |
SD, total RNA, heavy fraction, gPAR-CLIP, Replicate 2 |
| GSM1186346 |
SD, polyA RNA, light fraction, gPAR-CLIP, Replicate 1 |
| GSM1186347 |
SD, polyA RNA, light fraction, gPAR-CLIP, Replicate 2 |
| GSM1186348 |
SD-glucose, polyA RNA, light fraction, gPAR-CLIP, Replicate 1 |
| GSM1186349 |
SD-glucose, polyA RNA, light fraction, gPAR-CLIP, Replicate 2 |
| GSM1186350 |
SD-nitrogen polyA RNA, light fraction, gPAR-CLIP, Replicate 1 |
| GSM1186351 |
SD-nitrogen polyA RNA, light fraction, gPAR-CLIP, Replicate 2 |
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| Relations |
| BioProject |
PRJNA212138 |
| SRA |
SRP027402 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE48888_glucose_polyA_light_gPAR-CLIP_cluster.bed.gz |
298.5 Kb |
(ftp)(http) |
BED |
| GSE48888_nitrogen_polyA_light_gPAR-CLIP_cluster.bed.gz |
348.5 Kb |
(ftp)(http) |
BED |
| GSE48888_polyA_light_gPAR-CLIP_cluster.bed.gz |
538.1 Kb |
(ftp)(http) |
BED |
| GSE48888_total_heavy_gPAR-CLIP_cluster.bed.gz |
989 b |
(ftp)(http) |
BED |
| GSE48888_total_light_gPAR-CLIP_cluster.bed.gz |
461.2 Kb |
(ftp)(http) |
BED |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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