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| Status |
Public on Apr 17, 2020 |
| Title |
Prp5-Spt8/Spt3 Interaction Mediates a Reciprocal Coupling between Splicing and Transcription |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
Transcription and pre-mRNA splicing are coupled to promote gene expression and regulation. However, mechanisms by which transcription and splicing influence each other are still under investigation. The ATPase Prp5p is required for pre-spliceosome assembly and splicing proofreading at the branch-point region. From an open UV mutagenesis screen for genetic suppressors of prp5 defects and subsequent targeted testing, we identify components of the TBP-binding module of the Spt–Ada–Gcn5 Acetyltransferase (SAGA) complex, Spt8p and Spt3p. Spt8Δ and spt3Δ rescue the cold-sensitivity of prp5-GAR allele, and prp5 mutants restore growth of spt8Δ and spt3Δ strains on 6-azauracil. By chromatin immunoprecipitation (ChIP), we find that prp5 alleles decrease recruitment of RNA polymerase II (Pol II) to an intron-containing gene, which is rescued by spt8Δ. Further ChIP-seq reveals that global effects on Pol II-binding is mutually rescued by prp5 mutant and spt8Δ. Inhibited splicing caused by prp5-GAR is also restored by spt8Δ. In vitro assay indicates that Prp5p directly interacts with Spt8p, but not Spt3p. We demonstrate that Prp5p’s splicing proofreading is modulated by Spt8p and Spt3p. Therefore, this study reveals that interactions between the TBP-binding module of SAGA and the spliceosomal ATPase Prp5p mediate a balance between transcription initiation/elongation and pre-spliceosome assembly.
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| Overall design |
Examining the Pol II profile in WT and Spt8Δ strains with Prp5-WT or Prp5-GAR mutation at 16°C and 30°C
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| Contributor(s) |
Shao W, Ding Z, Query CC, Xu Y |
| Citation(s) |
32399566 |
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| Submission date |
Mar 18, 2020 |
| Last update date |
Jun 25, 2020 |
| Contact name |
Zhan Ding |
| E-mail(s) |
dingzhan@cemps.ac.cn
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| Organization name |
CAS Center for Excellence in Molecular Plant Science
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| Street address |
FengLin 300
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| City |
Shanghai |
| ZIP/Postal code |
200032 |
| Country |
China |
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| Platforms (1) |
| GPL26171 |
HiSeq X Ten (Saccharomyces cerevisiae) |
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| Samples (24)
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| GSM4435312 |
WT,GAR--30C-input |
| GSM4435313 |
WT,GAR--30C-IP rep1 |
| GSM4435314 |
WT,GAR--30C-IP rep2 |
| GSM4435315 |
WT,WT--16C-input |
| GSM4435316 |
WT,WT--16C-IP rep1 |
| GSM4435317 |
WT,WT--16C-IP rep2 |
| GSM4435318 |
WT,WT--30C-input |
| GSM4435319 |
WT,WT--30C-IP rep1 |
| GSM4435320 |
WT,WT--30C-IP rep2 |
| GSM4435321 |
spt8-ko,GAR--16C-input |
| GSM4435322 |
spt8-ko,GAR--16C-IP rep1 |
| GSM4435323 |
spt8-ko,GAR--16C-IP rep2 |
| GSM4435324 |
spt8-ko,GAR--30C-input |
| GSM4435325 |
spt8-ko,GAR--30C-IP rep1 |
| GSM4435326 |
spt8-ko,GAR--30C-IP rep2 |
| GSM4435327 |
spt8-ko,WT--16C-input |
| GSM4435328 |
spt8-ko,WT--16C-IP rep1 |
| GSM4435329 |
spt8-ko,WT--16C-IP rep2 |
| GSM4435330 |
spt8-ko,WT--30C-input |
| GSM4435331 |
spt8-ko,WT--30C-IP rep1 |
| GSM4435332 |
spt8-ko,WT--30C-IP rep2 |
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| Relations |
| BioProject |
PRJNA613270 |
| SRA |
SRP253236 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE147179_RAW.tar |
537.5 Mb |
(http)(custom) |
TAR (of BIGWIG) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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