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| Status |
Public on Sep 27, 2011 |
| Title |
Sub1 and RPA associate with RNA Polymerase II at different stages of transcription |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Genome binding/occupancy profiling by genome tiling array
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| Summary |
Single stranded DNA binding proteins play many roles in nucleic acid metabolism, but their importance during transcription remains unclear. Quantitative proteomic analysis of RNA polymerase II (RNApII) pre-initiation complexes (PICs) identified Sub1 and the Replication Protein A complex (RPA), both of which bind single-stranded DNA (ssDNA). Sub1, homolog of mammalian coactivator PC4, exhibits strong genetic interactions with factors necessary for promoter melting. Sub1 localizes near the transcription bubble in vitro and binds to promoters in vivo dependent upon PIC assembly. In contrast, RPA localizes to transcribed regions of active genes, strongly correlated with transcribing RNApII but independently of replication. RFA1 interacts genetically with transcription elongation factor genes. Interestingly, RPA levels increase at active promoters in cells carrying a Sub1 deletion or ssDNA binding mutant, suggesting competition for a common binding site. We propose that Sub1 and RPA interact with the non-template strand of RNApII complexes during initiation and elongation, respectively.
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| Overall design |
Chip-chip from wt and sub1D cells with Rfa1
Chromatin immunoprecipitation (ChIP) of Rfa1 in wt and sub1D yeast demonstrated that Rfa1 localization correlates with RNA Polymerase II and is increased at some transcription start sites when Sub1 has been deleted.
Comparison of Rfa1 localization in wt vs sub1D yeast
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| Contributor(s) |
Sikorski T, Holik J, Rando O, Buratowski S |
| Citation(s) |
22055186 |
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| Submission date |
Sep 27, 2011 |
| Last update date |
Sep 07, 2013 |
| Contact name |
Oliver Rando |
| E-mail(s) |
[email protected]
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| Phone |
508-856-8879
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| Organization name |
UMass Medical School
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| Street address |
364 Plantation St.
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| City |
Worcester |
| State/province |
MA |
| ZIP/Postal code |
01605-4321 |
| Country |
USA |
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| Platforms (1) |
| GPL10930 |
Agilent-014810 Yeast Whole Genome ChIP-on-Chip Microarray 4x44K (G4493A) [Probe Name version] |
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| Samples (4)
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| GSM802435 |
Rfa1 ChIP from wild-type cells, replicate 1 |
| GSM802436 |
Rfa1 ChIP from wild-type cells, replicate 2 |
| GSM802437 |
Rfa1 ChIP from sub1D cells, replicate 1 |
| GSM802438 |
Rfa1 ChIP from sub1D cells, replicate 2 |
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| Relations |
| BioProject |
PRJNA147897 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE32416_RAW.tar |
23.6 Mb |
(http)(custom) |
TAR (of GPR) |
| Processed data included within Sample table |
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