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| Status |
Public on Apr 02, 2016 |
| Title |
Localization of RNAPII and 3’ end formation factor CstF subunits on C. elegans genes and operons [ChIP-seq] |
| Organism |
Caenorhabditis elegans |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
Transcription termination is mechanistically coupled to pre-mRNA 3’ end formation to prevent transcription much beyond the gene 3’ end. C. elegans, however, engages in polycistronic transcription of operons in which 3’ end formation between genes is not accompanied by termination. We have performed RNA polymerase II (RNAPII) and CstF ChIP-seq experiments to investigate at a genome-wide level how RNAPII can transcribe through multiple poly-A signals without causing termination. Our data shows that transcription proceeds in some ways as if operons were composed of multiple adjacent single genes. Total RNAPII shows a small peak at the promoter of the gene cluster and a much larger peak at 3’ ends. These 3’ peaks coincide with maximal phosphorylation of Ser2 within the C-terminal domain (CTD) of RNAPII and maximal localization of the 3’ end formation factor CstF. This pattern occurs at all 3’ ends including those at internal sites in operons where termination does not occur. Thus the normal mechanism of 3’ end formation does not always result in transcription termination. Furthermore, reduction of CstF50 by RNAi did not substantially alter the pattern of CstF64, total RNAPII, or Ser2 phosphorylation at either internal or terminal 3’ ends. However, CstF50 RNAi did result in a subtle reduction of CstF64 binding upstream of the site of 3’ cleavage, suggesting that the CstF50/CTD interaction may facilitate bringing the 3’ end machinery to the transcription complex.
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| Overall design |
ChIP-seq examination of RNAPII and CstF subunits using C. elegans in duplicates.
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| Contributor(s) |
Blumenthal T, Saldi T |
| Citation missing |
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| Submission date |
Apr 01, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Alfonso Garrido-Lecca |
| E-mail(s) |
alfonso.garrido-lecca@colorado.edu
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| Organization name |
CU Boulder
|
| Street address |
347 UCB
|
| City |
Boulder |
| ZIP/Postal code |
80309 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL13657 |
Illumina HiSeq 2000 (Caenorhabditis elegans) |
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| Samples (18)
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| This SubSeries is part of SuperSeries: |
| GSE79823 |
Localization of RNAPII and 3 end formation factor CstF subunits on C. elegans genes and operons |
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| Relations |
| BioProject |
PRJNA317071 |
| SRA |
SRP072749 |
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