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| Status |
Public on Oct 01, 2019 |
| Title |
Chromatin-dependent cryptic promoters encode alternative protein isoforms in yeast |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Cryptic transcription is widespread and generates a heterogeneous group of RNA molecules of unknown function. To improve our understanding of cryptic transcription, we investigated their transcription start site usage, chromatin organization and post-transcriptional consequences in Saccharomyces cerevisiae. We show that transcription start sites of chromatin-dependent internal cryptic transcripts resemble those of protein coding genes in terms of DNA sequence, directionality and chromatin accessibility. We define the 5’ and 3’ boundaries of cryptic transcripts and show that, contrary to RNA degradation-dependent ones , they often overlap with the end of the gene thereby using the canonical polyadenylation site and associate to polyribosomes. In fact, we show that chromatin-dependent cryptic transcripts can be recognized by ribosomes and may produce truncated polypeptides from downstream, in-frame start codons. Our work suggests that a significant fraction of chromatin-dependent internal cryptic promoters are in fact alternative truncated mRNA isoforms. The expression of these chromatin-dependent isoforms is conserved from yeast to human expanding the functional consequences of cryptic transcription and proteome complexity.
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| Overall design |
We identify 5' transcription start sites applying our 5'cap -sequencing approach as previously described (Pelechano et al. Nat Protocols. 2016 PMID: 26820793). To measue polyribosome association we applied the 5'cap -sequencing approach to either total RNA extracts or polyribosome-associated fraction after ultracentrifugation in sucrose gradient. We performed 2 biologically independent experiments.
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| Contributor(s) |
Wei W, Hennig BP, Wang J, Zhang Y, Chabbert CD, Adjalley SH, Steinmetz LM, Pelechano V |
| Citation(s) |
31740578 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| P01 HG000205 |
Revolutionizing Biomedical and Clinical Research through Innovative Technology |
STANFORD UNIVERSITY |
Lars M Steinmetz |
|
| Submission date |
Aug 28, 2018 |
| Last update date |
Jan 02, 2020 |
| Contact name |
Vicent Pelechano |
| E-mail(s) |
vicente.pelechano.garcia@ki.se
|
| Organization name |
ScilifeLab - Karolinska Institutet
|
| Department |
MTC
|
| Street address |
Nobels väg 16
|
| City |
Solna |
| ZIP/Postal code |
SE-17177 |
| Country |
Sweden |
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| Platforms (1) |
| GPL13821 |
Illumina HiSeq 2000 (Saccharomyces cerevisiae) |
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| Samples (16)
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| Relations |
| BioProject |
PRJNA488219 |
| SRA |
SRP159001 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE119114_BY4741.normalized.minus.bedGraph.gz |
4.5 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_BY4741.normalized.plus.bedGraph.gz |
4.5 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_by_pol1_UMI_5end_raw.bedGraph.gz |
7.3 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_by_pol2_UMI_5end_raw.bedGraph.gz |
9.1 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_by_tot1_UMI_5end_raw.bedGraph.gz |
11.6 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_by_tot2_UMI_5end_raw.bedGraph.gz |
8.0 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_eaf3.normalized.minus.bedGraph.gz |
4.8 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_eaf3.normalized.plus.bedGraph.gz |
4.9 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_rco1.normalized.minus.bedGraph.gz |
5.7 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_rco1.normalized.plus.bedGraph.gz |
5.8 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_rrp6.normalized.minus.bedGraph.gz |
6.3 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_rrp6.normalized.plus.bedGraph.gz |
6.3 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_set1.normalized.minus.bedGraph.gz |
4.0 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_set1.normalized.plus.bedGraph.gz |
4.0 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_set2.normalized.minus.bedGraph.gz |
6.3 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE119114_set2.normalized.plus.bedGraph.gz |
6.4 Mb |
(ftp)(http) |
BEDGRAPH |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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