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| Status |
Public on Mar 29, 2019 |
| Title |
Slx5/Slx8-dependent ubiquitin hotspots on chromatin contribute to stress tolerance [ChIP-chip] |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Genome binding/occupancy profiling by genome tiling array
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| Summary |
Chromatin is a highly regulated environment, and protein association with chromatin is often controlled by post-translational modifications and the corresponding enzymatic machinery. Specifically, SUMO-targeted ubiquitin ligases (STUbLs) have emerged as key players in nuclear quality control, genome maintenance and transcription. However, how STUbLs select specific substrates amongst myriads of SUMOylated proteins on chromatin remains unclear. Here, we reveal a remarkable co-localization of the budding yeast STUbL Slx5/Slx8 and ubiquitin at seven genomic loci that we term ‘ubiquitin hotspots’. Ubiquitylation at these sites depends on Slx5/Slx8 and protein turnover on the Cdc48 segregase. We identify the transcription factor-like Ymr111c/Euc1 to associate with these sites and be a critical determinant of ubiquitylation. Euc1 specifically targets Slx5/Slx8 to ubiquitin hotspots via bipartite binding of Slx5 that involves the Slx5 SUMO-interacting motifs and an additional, novel substrate recognition domain. Interestingly, the Euc1-ubiquitin hotspot pathway acts redundantly with chromatin modifiers of the H2A.Z and Rpd3L pathways in specific stress responses. Thus, our data suggest that STUbL-dependent ubiquitin hotspots shape chromatin during stress adaptation.
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| Overall design |
ChIP-chip profiling of ubiquitylated proteins (FK2-antibody and K48-specific Apu2-clone), Slx8-9myc and Ymr111c/Euc1 in S. cerevisiae. Custom-made Nimblegen arrays (100304_Scer2_MS_Chip_Stefan Jentsch MPI Biochemistry S.cerevisiae 135K Tiling Array Version 2) were used to compare ChIP versus input DNA. Typically, replicate experiments were performed including a dye-swap for ChIP/input DNA (except for rad6-delta-and one of the IgG-control experiments in Fig. 1A).
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| Contributor(s) |
Höpfler M, Kern MJ, Straub T, Prytuliak R, Habermann BH, Pfander B, Jentsch S |
| Citation(s) |
31015336 |
| |
| Submission date |
Aug 20, 2018 |
| Last update date |
Jun 28, 2019 |
| Contact name |
Markus Höpfler |
| E-mail(s) |
hoepfler@biochem.mpg.de
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| Organization name |
Max-Planck Institute of Biochemistry
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| Department |
Molecular Cell Biology
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| Lab |
Stefan Jentsch
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| Street address |
Am Klopferspitz 18
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| City |
Martinsried/Munich |
| ZIP/Postal code |
82152 |
| Country |
Germany |
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| Platforms (1) |
| GPL16746 |
100304 Scer2 MS Chip Stefan Jentsch MPI Biochemistry S.cerevisiae 135K Tiling Array Version 2 |
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| Samples (23)
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| This SubSeries is part of SuperSeries: |
| GSE118818 |
Slx5/Slx8-dependent ubiquitin hotspots on chromatin contribute to stress tolerance |
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| Relations |
| BioProject |
PRJNA486777 |
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