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Series GSE113835 Query DataSets for GSE113835
Status Public on Sep 10, 2019
Title SUMO-chain-regulated proteasomal degradation timing exemplified in DNA replication initiation
Organism Saccharomyces cerevisiae
Experiment type Genome binding/occupancy profiling by genome tiling array
Summary Similar to ubiquitin, SUMO forms chains, but the identity of SUMO-chain-modified factors and the purpose of this modification remain largely unknown. Here, we identify budding yeast SUMO protease Ulp2, able to disassemble SUMO chains, as a DDK interactor enriched at replication origins that promotes DNA replication initiation. Replication-engaged DDK is SUMOylated on chromatin, becoming a degradation-prone substrate when Ulp2 no longer protects it against SUMO-chain assembly. Specifically, SUMO chains channel DDK for SUMO-targeted ubiquitin ligase Slx5/Slx8-mediated and Cdc48 segregase-assisted proteasomal degradation. Importantly, the SUMOylation-defective ddk-KR mutant rescues inefficient replication onset and MCM activation in cells lacking Ulp2, suggesting that SUMO chains time DDK degradation. Using two unbiased proteomic approaches, we further identify subunits of the MCM helicase and other factors as SUMO-chain-modified degradation-prone substrates of Ulp2 and Slx5/Slx8. We thus propose SUMO-chain-/Ulp2-protease-regulated proteasomal degradation as a mechanism that times the availability of functionally-engaged SUMO-modified protein pools during replication and beyond.
 
Overall design Protein binding profiles by ChIP-on-chip analysis were generated as described (Bermejo et al., 2009). BrdU incorporation profiles by BrdU IP-on-chip have been generated as described (Katou et al., 2003). Labeled probes were hybridized to Affymetrix S.cerevisiae Tiling 1.0 (P/N 900645) arrays and processed with TAS software.
 
Contributor(s) Psakhye I, Castellucci F, Branzei D
Citation(s) 31519521
Submission date Apr 30, 2018
Last update date Dec 13, 2019
Contact name Ivan Psakhye
Organization name IFOM, the FIRC Institute of Molecular Oncology
Lab DNA Repair
Street address Via Adamello 16
City Milan
ZIP/Postal code 20139
Country Italy
 
Platforms (1)
GPL7250 [Sc03b_MR] Affymetrix GeneChip S. cerevisiae Tiling 1.0R Array
Samples (19)
GSM3120927 Ulp2-PK G1
GSM3120928 Ulp2-PK HU
GSM3120929 Ulp2-FLAG HU
Relations
BioProject PRJNA454214

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Supplementary file Size Download File type/resource
GSE113835_RAW.tar 1.7 Gb (http)(custom) TAR (of BAR, CEL)
Processed data provided as supplementary file

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