Dual roles of the SUMO-interacting motif in the regulation of Srs2 sumoylation

Kolesar, Peter; Sarangi, Prabha; Altmannová, Veronika; Zhao, Xiaolan; Krejčí, Lumír

doi:10.1093/nar/gks484

Cited by 58 publications

(90 citation statements)

References 70 publications

(83 reference statements)

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“…Compared with the 0 hr of mitosis time point, an approximate fivefold increase in Srs2 was seen at 4-8 hr of meiosis. Overexpressed Srs2 generated a ladder on Western blotting, suggesting multiple post-translational modifications, e.g., SUMOlyation (Kolesar et al 2012).…”

Section: Meiotic Overexpression Of Srs2 Results In Dosage-dependent Tmentioning

confidence: 99%

Remodeling of the Rad51 DNA Strand-Exchange Protein by the Srs2 Helicase

2013

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Homologous recombination is associated with the dynamic assembly and disassembly of DNA-protein complexes. Assembly of a nucleoprotein filament comprising ssDNA and the RecA homolog, Rad51, is a key step required for homology search during recombination. The budding yeast Srs2 DNA translocase is known to dismantle Rad51 filament in vitro. However, there is limited evidence to support the dismantling activity of Srs2 in vivo. Here, we show that Srs2 indeed disrupts Rad51-containing complexes from chromosomes during meiosis. Overexpression of Srs2 during the meiotic prophase impairs meiotic recombination and removes Rad51 from meiotic chromosomes. This dismantling activity is specific for Rad51, as Srs2 Overexpression does not remove Dmc1 (a meiosis-specific Rad51 homolog), Rad52 (a Rad51 mediator), or replication protein A (RPA; a single-stranded DNA-binding protein). Rather, RPA replaces Rad51 under these conditions. A mutant Srs2 lacking helicase activity cannot remove Rad51 from meiotic chromosomes. Interestingly, the Rad51-binding domain of Srs2, which is critical for Rad51-dismantling activity in vitro, is not essential for this activity in vivo. Our results suggest that a precise level of Srs2, in the form of the Srs2 translocase, is required to appropriately regulate the Rad51 nucleoprotein filament dynamics during meiosis. RECOMBINATION between homologous sequences promotes genomic stability. As such, a wide variety of DNAassociated proteins and protein complexes regulate the recombination process. Within chromatin, the chromosome structure probably also directly affects the assembly and disassembly of protein complexes involved in recombination. To understand the molecular mechanisms that govern assembly and disassembly of these complexes, homologous recombination during meiosis can be used as a model system.Meiotic recombination is initiated when Spo11, a meiosisspecific topoisomerase-like protein, generates double-strand breaks (DSBs) (Keeney 2001). The 59 ends of these DSBs are quickly resected to produce a 39-overhanging stretch of single-stranded DNA (ssDNA). The ssDNA is used to search for homologous double-stranded DNAs (dsDNAs). Once homology between the ss-and dsDNAs are matched, the ssDNA is invaded into the dsDNA. Strand invasion leads to DNA synthesis with the 39 end of the invading strand as the primer. The resulting intermediate is then converted into the singleinvasion intermediate (SEI) and then a DNA structure with double Holiday junctions (dHJ) (Schwacha and Kleckner 1994;Hunter and Kleckner 2001). Resolution of dHJ structures typically results in chromosomal crossover (CO) (Allers and Lichten 2001;Hunter and Kleckner 2001). Alternatively, newly synthesized DNA of the invading strand after the displacement can anneal with ssDNA from the other end of the DSB, which results in the formation of a noncrossover (NCO) product (through the synthesis-dependent strand-annealing pathway). In meiosis, the recombination occurs preferentially between homologous chromosomes rather than betw...

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Section: Meiotic Overexpression Of Srs2 Results In Dosage-dependent Tmentioning

confidence: 99%

Remodeling of the Rad51 DNA Strand-Exchange Protein by the Srs2 Helicase

2013

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“…jugation to a SUMO conjugation site (25,26). In this case mutagenesis of amino acids Ile 388 and Leu 391 in FXR to alanine (I388A and L391A), which in other studies was sufficient to disrupt the SIM, had no effect on in vivo SUMOylation of the mutant FXR (Fig.…”

Section: Methodsmentioning

confidence: 91%

“…FXR in fact has a SIM consensus site. There are now multiple proteins for which SUMOylation is dependent on the presence of a SIM in the substrate (25,26). In this scenario SUMO binding to the SIM domain would be the initial association with its target protein and would precede SUMO conjugation to the SUMO consensus motif.…”

Section: Discussionmentioning

confidence: 99%

SUMOylation of the Farnesoid X Receptor (FXR) Regulates the Expression of FXR Target Genes

Balasubramaniyan

Luo

Sun

et al. 2013

Journal of Biological Chemistry

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Background: Small ubiquitin-like modifiers (SUMO) are covalently conjugated to other proteins including nuclear receptors leading to modification of various cellular processes. Results: Ligand-dependent SUMOylation of farnesoid X receptor (FXR) negatively regulates the expression of its target genes. Conclusion: SUMO modification attenuates the capacity of FXR to function as a transcriptional activator. Significance: Defining post-translation modification of FXR by SUMO is important to understanding how this nuclear receptor functions in health and disease.

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“…This explains the srs2 Δ puzzling phenotype which includes both recombination deficiency and hyper‐recombination. The anti‐recombination role of Srs2 is particularly important for inhibition of recombination at replication forks and involves complex regulation of Srs2 through post‐translational modifications at its C‐terminus (Saponaro et al , 2010; Kolesar et al , 2012). The C‐terminal part is not needed for the role of Srs2 in re‐synthesis of resected DNA, and therefore, the Srs2 pro‐recombination function is genetically separable from its role at replication forks.…”

Section: Discussionmentioning

confidence: 99%

Unloading of homologous recombination factors is required for restoring double‐stranded DNA at damage repair loci

et al. 2017

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Cells use homology‐dependent DNA repair to mend chromosome breaks and restore broken replication forks, thereby ensuring genome stability and cell survival. DNA break repair via homology‐based mechanisms involves nuclease‐dependent DNA end resection, which generates long tracts of single‐stranded DNA required for checkpoint activation and loading of homologous recombination proteins Rad52/51/55/57. While recruitment of the homologous recombination machinery is well characterized, it is not known how its presence at repair loci is coordinated with downstream re‐synthesis of resected DNA. We show that Rad51 inhibits recruitment of proliferating cell nuclear antigen (PCNA), the platform for assembly of the DNA replication machinery, and that unloading of Rad51 by Srs2 helicase is required for efficient PCNA loading and restoration of resected DNA. As a result, srs2Δ mutants are deficient in DNA repair correlating with extensive DNA processing, but this defect in srs2Δ mutants can be suppressed by inactivation of the resection nuclease Exo1. We propose a model in which during re‐synthesis of resected DNA, the replication machinery must catch up with the preceding processing nucleases, in order to close the single‐stranded gap and terminate further resection.

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Dual roles of the SUMO-interacting motif in the regulation of Srs2 sumoylation

Cited by 58 publications

References 70 publications

Remodeling of the Rad51 DNA Strand-Exchange Protein by the Srs2 Helicase

Remodeling of the Rad51 DNA Strand-Exchange Protein by the Srs2 Helicase

SUMOylation of the Farnesoid X Receptor (FXR) Regulates the Expression of FXR Target Genes

Unloading of homologous recombination factors is required for restoring double‐stranded DNA at damage repair loci

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