Ubc9 Sumoylation Controls SUMO Chain Formation and Meiotic Synapsis in Saccharomyces cerevisiae

Klug, Helene; Xaver, Martin; Chaugule, Viduth K.; Koidl, Stefanie; Mittler, Gerhard; Klein, Franz; Pichler, Andrea

doi:10.1016/j.molcel.2013.03.027

Cited by 67 publications

(74 citation statements)

References 35 publications

(57 reference statements)

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“…Nevertheless, polysumoylation is important for specific processes, such as meiosis (Cheng et al 2006;Klug et al 2013) and maintenance of higher-order chromatin structure (Srikumar et al 2013a). A difficulty in studying aspects of polySUMO function in yeast has been the lack of effective affinity reagents for isolating polySUMO-modified proteins.…”

Section: Discussionmentioning

confidence: 99%

SUMO Pathway Modulation of Regulatory Protein Binding at the Ribosomal DNA Locus inSaccharomyces cerevisiae

Gillies

Hickey

et al. 2016

Genetics

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In this report, we identify cellular targets of Ulp2, one of two Saccharomyces cerevisiae small ubiquitin-related modifier (SUMO) proteases, and investigate the function of SUMO modification of these proteins. PolySUMO conjugates from ulp2D and ulp2D slx5D cells were isolated using an engineered affinity reagent containing the four SUMO-interacting motifs (SIMs) of Slx5, a component of the Slx5/Slx8 SUMO-targeted ubiquitin ligase (STUbL). Two proteins identified, Net1 and Tof2, regulate ribosomal DNA (rDNA) silencing and were found to be hypersumoylated in ulp2D, slx5D, and ulp2D slx5D cells. The increase in sumoylation of Net1 and Tof2 in ulp2D, but not ulp1ts cells, indicates that these nucleolar proteins are specific substrates of Ulp2. Based on quantitative chromatin-immunoprecipitation assays, both Net1 and Tof2 lose binding to their rDNA sites in ulp2D cells and both factors largely regain this association in ulp2D slx5D. A parsimonious interpretation of these results is that hypersumoylation of these proteins causes them to be ubiquitylated by Slx5/ Slx8, impairing their association with rDNA. Fob1, a protein that anchors both Net1 and Tof2 to the replication-fork barrier (RFB) in the rDNA repeats, is sumoylated in wild-type cells, and its modification levels increase specifically in ulp2D cells. Fob1 experiences a 50% reduction in rDNA binding in ulp2D cells, which is also rescued by elimination of Slx5. Additionally, overexpression of Sir2, another RFBassociated factor, suppresses the growth defect of ulp2D cells. Our data suggest that regulation of rDNA regulatory proteins by Ulp2 and the Slx5/Slx8 STUbL may be the cause of multiple ulp2D cellular defects. KEYWORDS SUMO; Ulp2; rDNA; RENT complex; Fob1 P OST-TRANSLATIONAL protein modifications provide a common mechanism for regulating protein-protein interactions, protein localization, and protein degradation. The small ubiquitin-related modifier (SUMO) protein family modifies target proteins through covalent attachment to lysine side chains (Johnson 2004). The function of substrate "sumoylation" varies with the protein that is sumoylated, but a common role is to modulate the assembly of large protein complexes either by creating additional binding sites or by blocking existing sites (Kerscher 2007).Protein sumoylation occurs through an enzymatic cascade similar to ubiquitylation (Johnson 2004). In Saccharomyces cerevisiae, a single gene, SMT3, codes for SUMO, and as in most species, SUMO ligation is essential for viability. Removal of SUMO from target proteins is also an important regulatory step. S. cerevisiae has two SUMO proteases, Ulp1 and Ulp2 (Hickey et al. 2012). Most Ulp1 is bound to the nuclear pore complex (NPC), and the enzyme is essential for cell-cycle progression. NPC tethering regulates Ulp1 activity by restricting its access to sumoylated proteins (Li and Hochstrasser 2003;Panse et al. 2003). Ulp1 also processes the SUMO precursor by removing its last three residues, thereby exposing the C-terminal Gly-Gly motif required for SU...

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Section: Discussionmentioning

confidence: 99%

SUMO Pathway Modulation of Regulatory Protein Binding at the Ribosomal DNA Locus inSaccharomyces cerevisiae

Gillies

Hickey

et al. 2016

Genetics

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“…SUMOylation is a prominent feature of meiotic chromosomes. SUMO occurs along the SCs in budding yeast and humans, suggesting a role in SC formation (Cheng et al 2006;Hooker and Roeder 2006;Brown et al 2008;Klug et al 2013;VoelkelMeiman et al 2013). We therefore investigated whether Hei10 plays any role for SUMO localization during Sordaria meiotic prophase.…”

Section: Hei10's Rxl Domain Is Required To Elicit T1 Foci and Constramentioning

confidence: 99%

E3 ligase Hei10: a multifaceted structure-based signaling molecule with roles within and beyond meiosis

et al. 2014

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Human enhancer of invasion-10 (Hei10) mediates meiotic recombination and also plays roles in cell proliferation. Here we explore Hei10's roles throughout the sexual cycle of the fungus Sordaria with respect to localization and effects of null, RING-binding, and putative cyclin-binding (RXL) domain mutations. Hei10 makes three successive types of foci. Early foci form along synaptonemal complex (SC) central regions. At some of these positions, depending on its RING and RXL domains, Hei10 mediates development and turnover of two sequential types of recombination complexes, each demarked by characteristic amplified Hei10 foci. Integration with ultrastructural data for recombination nodules further reveals that recombination complexes differentiate into three types, one of which corresponds to crossover recombination events during or prior to SC formation. Finally, Hei10 positively and negatively modulates SUMO localization along SCs by its RING and RXL domains, respectively. The presented findings suggest that Hei10 integrates signals from the SC, associated recombination complexes, and the cell cycle to mediate both the development and programmed turnover/evolution of recombination complexes via SUMOylation/ubiquitination. Analogous cell cycle-linked assembly/disassembly switching could underlie localization and roles for Hei10 in centrosome/spindle pole body dynamics and associated nuclear trafficking. We suggest that Hei10 is a unique type of structure-based signal transduction protein.

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“…To test whether the cleavage of linear Smt3 chains by Ulp2 reflects its action on lysine-linked chains, we employed an in vivo sumoylation system reconstituted in E. coli to obtain a substrate with authentic Lys-linked poly-Smt3 chains (61). The SUMO-conjugating enzyme Ubc9 catalyzes its own sumoylation both in yeast cells (24) and in the E. coli-based system. This enabled the generation of poly-sumoylated Ubc9 (Fig.…”

Section: Ulp2 Processes Poly-smt3 Chains In a Sequentialmentioning

confidence: 99%

In Vitro Studies Reveal a Sequential Mode of Chain Processing by the Yeast SUMO (Small Ubiquitin-related Modifier)-specific Protease Ulp2

Eckhoff¹,

Dohmen

2015

Journal of Biological Chemistry

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Background: Modification of proteins with small ubiquitin-related modifier has many important functions in cell biology. Results: Substrate-attached SUMO chains are processed by the yeast enzymes Ulp1 and Ulp2 via different mechanisms. Conclusion: Ulp2 recognizes chains with three or more SUMO units and releases single SUMO molecules from their distal ends. Significance: Ulp2 controls the dynamic length range of cellular SUMO chains.

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Ubc9 Sumoylation Controls SUMO Chain Formation and Meiotic Synapsis in Saccharomyces cerevisiae

Cited by 67 publications

References 35 publications

SUMO Pathway Modulation of Regulatory Protein Binding at the Ribosomal DNA Locus inSaccharomyces cerevisiae

SUMO Pathway Modulation of Regulatory Protein Binding at the Ribosomal DNA Locus inSaccharomyces cerevisiae

E3 ligase Hei10: a multifaceted structure-based signaling molecule with roles within and beyond meiosis

In Vitro Studies Reveal a Sequential Mode of Chain Processing by the Yeast SUMO (Small Ubiquitin-related Modifier)-specific Protease Ulp2

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