Mus81-Eme1 Are Essential Components of a Holliday Junction Resolvase

Boddy, Michael N.; Gaillard, Patrick; McDonald, W. Hayes; Shanahan, Paul; Yates, John R.; Russell, Paul

doi:10.1016/s0092-8674(01)00536-0

Cited by 503 publications

(545 citation statements)

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“…The nse1-1 meiotic phenotype suggests that the normal meiotic nuclear divisions are blocked. This phenotype is reminiscent of that observed in the meiotic recombination-defective mutant mus81 (Boddy et al, 2001). In this mutant, DNA fails to segregate at meiosis I due to unresolved recombination structures.…”

Section: Nse1 Nse2 and Nse3 Play A Critical Role In Meiosismentioning

confidence: 69%

“…For example, budding yeast Rad54 mutants are DSB repair defective and arrest in meiotic prophase, whereas fission yeast lacking the Rad54 homologue Rhp54, continue through meiosis despite unrepaired DSBs (Shinohara et al, 1997;Catlett and Forsburg, 2003). In addition, Rhp51 mutants were shown to complete meiosis, despite persistent DSBs (Zenvirth and Simchen, 2000;Boddy et al, 2001). In both Rad54 and Rhp51 mutants DNA segregates remarkably well, with asci often containing four spores, each of which has DNA in it.…”

Section: Discussionmentioning

confidence: 99%

“…Cells were photographed with Nikon Eclipse E800 microscope equipped with a Photometrics Quantix charge-coupled device camera. Samples for visualization of meiotic figures were prepared as described previously (Boddy et al, 2001). …”

Section: Immunoblotting and Microscopy Techniquesmentioning

confidence: 99%

“…Proteins associating with Smc5-TAP were identified by multidimensional protein identification technology (MudPIT) by using established methods (Boddy et al, 2001;Washburn et al, 2001;McDonald and Yates, 2002). Briefly, cells (ϳ50 g wet weight) expressing Smc5-TAP at the genomic locus were frozen in liquid nitrogen and lysed using a motorized mortar and pestle (Retsch, Newtown, PA) in buffer A (50 mM Tris pH 8, 150 mM NaCl, 2 mM EDTA, 10% glycerol, 0.2% Nonidet P-40, 5 g/ml each of leupeptin, pepstatin, and aprotinin, and 1 mM PMSF).…”

Section: Identification Of Smc5 Interacting Proteinsmentioning

confidence: 99%

“…Proteins in the eluate were precipitated and subjected to mass spectrometry analysis. We have used this system extensively for identification of protein complexes in S. pombe and have thus generated a comprehensive list of common background proteins (Boddy et al, 2001;Boddy et al, 2003;McDonald et al, 2003). Comparison of the Smc5-TAP data set with previous purifications allowed us to exclude common background such as ribosomal proteins, abundant metabolic enzymes, and actin.…”

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confidence: 99%

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Nse1, Nse2, and a Novel Subunit of the Smc5-Smc6 Complex, Nse3, Play a Crucial Role in Meiosis

Pébernard

McDonald

Pavlova

et al. 2004

MBoC

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The structural maintenance of chromosomes (SMC) family of proteins play key roles in the organization, packaging, and repair of chromosomes. Cohesin (Smc1؉3) holds replicated sister chromatids together until mitosis, condensin (Smc2؉4) acts in chromosome condensation, and Smc5؉6 performs currently enigmatic roles in DNA repair and chromatin structure. The SMC heterodimers must associate with non-SMC subunits to perform their functions. Using both biochemical and genetic methods, we have isolated a novel subunit of the Smc5؉6 complex, Nse3. Nse3 is an essential nuclear protein that is required for normal mitotic chromosome segregation and cellular resistance to a number of genotoxic agents. Epistasis with Rhp51 (Rad51) suggests that like Smc5؉6, Nse3 functions in the homologous recombination based repair of DNA damage. We previously identified two non-SMC subunits of Smc5؉6 called Nse1 and Nse2. Analysis of nse1-1, nse2-1, and nse3-1 mutants demonstrates that they are crucial for meiosis. The Nse1 mutant displays meiotic DNA segregation and homologous recombination defects. Spore viability is reduced by nse2-1 and nse3-1, without affecting interhomolog recombination. Finally, genetic interactions shared by the nse mutants suggest that the Smc5؉6 complex is important for replication fork stability. INTRODUCTIONBoth endogenous and exogenous agents constantly threaten genomic integrity. The DNA double-strand break (DSB) is a potentially life-threatening lesion for a cell and can occur spontaneously during growth, as part of a programmed event such as meiosis or as the result of exposure to environmental genotoxic agents. Multiple mechanisms exist to repair DSBs, including nonhomologous end joining and homologous recombination (HR) (Paques and Haber, 1999;Symington, 2002). The HR pathway serves to maintain all original genetic information during DSB repair. Many components of that pathway have been identified and characterized (Paques and Haber, 1999;Symington, 2002). HR involves multiple steps. The DSB is first resected to generate a recombinogenic 3Ј overhang that invades a homologous sequence (e.g., sister chromatid), forming a displacement or D-loop. The invasion step is catalyzed by a number of proteins including the Escherichia coli RecA homologue, Rad51. D-loop formation primes repair synthesis, which is followed by resolution of the recombined duplexes and ligation, producing intact duplex DNA molecules (Paques and Haber, 1999;Symington, 2002). The resolution of recombined DNA duplexes can yield products in which there has been reciprocal exchange of flanking markers (crossover) or not. During mitotic growth gene conversion is infrequently accompanied by crossover, whereas during meiosis, crossover recombination is much more prevalent (Paques and Haber, 1999).Efficient DNA repair requires modulation of both local and higher order chromatin structure. Interestingly, the structural maintenance of chromosomes (SMC) proteins have recently been recognized as important players in DNA repair (Hirano, 2002;Jessberger...

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Section: Nse1 Nse2 and Nse3 Play A Critical Role In Meiosismentioning

confidence: 69%

Section: Discussionmentioning

confidence: 99%

Section: Immunoblotting and Microscopy Techniquesmentioning

confidence: 99%

Section: Identification Of Smc5 Interacting Proteinsmentioning

confidence: 99%

mentioning

confidence: 99%

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Nse1, Nse2, and a Novel Subunit of the Smc5-Smc6 Complex, Nse3, Play a Crucial Role in Meiosis

Pébernard

McDonald

Pavlova

et al. 2004

MBoC

Self Cite

112

150

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Holliday junction‐resolving enzymes—structures and mechanisms

Lilley

2017

FEBS Letters

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Holliday junction-resolving enzymes are nucleases that are highly specific for the structure of the junction, to which they bind in dimeric form. Two symmetrically disposed cleavages are made. These are not simultaneous, but the second cleavage is accelerated relative to the first, so ensuring that bilateral cleavage occurs during the lifetime of the DNA-protein complex. In eukaryotic cells there are two known junction-resolving activities. GEN1 is similar to enzymes from lower organisms. A crystallographic structure of a fungal GEN1 bound to the product of resolution has been determined. These complexes are dimerized within the crystal lattice such that the strands of the products may be simply reconnected to form a junction. These structures suggest a trajectory for the resolution process.

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Control of Mus81 nuclease during the cell cycle

Pfander

Matos

2017

FEBS Letters

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Edited by Wilhelm JustDNA replication and homologous recombination involve the formation of branched DNA structures that physically link chromosomes. Such DNAbased connections, which arise during S-phase, are typically disengaged prior to entry into mitosis, in order to ensure proper chromosome segregation. Exceptions can, however, occur: replication stress, or elevated levels of DNA damage, may cause cells to enter mitosis with unfinished replication as well as carrying recombination intermediates, such as Holliday junctions. Hence, cells are equipped with pathways that recognize and process branched DNA structures, and evolved mechanisms to enhance their function when on the verge of undergoing cell division. One of these pathways utilizes the structure-selective endonuclease Mus81, which is thought to facilitate the resolution of replication and recombination intermediates. Mus81 function is known to be enhanced upon entry into M phase in budding yeast and human cells. Based on recent findings, we discuss here an updated model of Mus81 control during the cell cycle.

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Mus81-Eme1 Are Essential Components of a Holliday Junction Resolvase

Cited by 503 publications

References 47 publications

Nse1, Nse2, and a Novel Subunit of the Smc5-Smc6 Complex, Nse3, Play a Crucial Role in Meiosis

Nse1, Nse2, and a Novel Subunit of the Smc5-Smc6 Complex, Nse3, Play a Crucial Role in Meiosis

Holliday junction‐resolving enzymes—structures and mechanisms

Control of Mus81 nuclease during the cell cycle

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