The Smc5–Smc6 complex has important roles in DNA repair, probably in part through its association with specific additional subunits that participate in DNA repair. Its binding to chromosomes along chromosome arms, but not at sites of DNA damage, depends on Scc2–Scc4 (Betts Lindroos et al., 2006). The association pattern of the Smc5–Smc6
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Feb 14, 2021 · loaders in a nucleotide-bound DNA gripping state revealed the presence of two DNA binding modules (Higashi et al., 2020; Shi et al., 2020). One is the DNA gripping interaction that forms as the cohesin loader, Scc2Mis4, clamps the DNA onto the engaged SMC ATPase heads (Figure 1A, left). Both Scc2Mis4 and the SMC heads
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Jan 16, 2014 · Because Mis4 Scc2 contains a putative leucine zipper, we investigated DNA binding of Mis4 Scc2 /Ssl3 Scc4. We detected concentration-dependent DNA binding of Mis4 Scc2 /Ssl3 Scc4 to double stranded DNA (dsDNA) (Fig. 1c,d and Extended Data Fig. 1c). Single stranded (ssDNA) was bound poorly, while a Y-fork DNA substrate, mimicking open DNA structures that might exist at some …
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Dec 01, 2013 · Mis4 Scc2 /Ssl3 Scc4 is a DNA binding protein. a, Purified Mis4 Scc2 /Ssl3 Scc4 complex (M4S3) and its Mis4 Scc2 subunit (M4) were analyzed by SDS-PAGE and Coomassie blue staining (CBB) or Western blotting using antibodies directed against Mis4 or its C-terminal HA epitope. b, glycerol gradient centrifugation of the Mis4 Scc2 /Ssl3 Scc4 complex, followed by SDS-PAGE and silver staining.
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Jan 01, 2018 · Scc2, with its stable binding partner Scc4, promotes cohesin loading whereas Pds5, with its substoichiometric binding partner Wapl, dissociates cohesin from chromatin [31 – 33]. If the molecular function of cohesin involves topological embrace, the ring must be opened for both DNA entrapment and release.
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The biochemical reconstitution of topological DNA binding by budding yeast cohesin yields surprises and opens opportunities to exploit experimental approaches developed in this model organism. The ring-shaped chromosomal cohesin complex holds sister chromatids together by topological embrace, a prerequisite for accurate chromosome segregation.
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Sep 10, 2018 · (A) Effect of ADP/AlFx on loader binding to cohesin using DNA-bead assay. WT cohesin and an equimolar amount of loader was assayed for binding to DNA beads in the presence or ATP or ADP/AlFx. Silver-stained gel to assess cohesin (Psm3, Psm1, Rad21/Psc3) and loader (Mis4, Ssl3) bound to …
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Nov 10, 2020 · Pairwise Synchrony and Correlations Depend on the Structure of the Population Code in Visual Cortex. Veronika Koren, Ariana R. Andrei, Ming Hu, Valentin Dragoi, Klaus Obermayer. Article 108367. Download PDF.
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Dec 01, 2015 · Wapl Promotes DNA Entry into the Cohesin Ring. A purified fission-yeast cohesin tetramer, consisting of a Psm1-Psm3 dimer, Rad21, and Psc3, topologically loads onto DNA in vitro in a reaction that is facilitated by the Mis4-Ssl3 cohesin loader (Murayama and Uhlmann, 2014).Two additional cohesin subunits, Pds5 and Wapl, have been implicated in regulating cohesin's chromosome …
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Jan 01, 2018 · Scc2, with its stable binding partner Scc4, promotes cohesin loading whereas Pds5, with its substoichiometric binding partner Wapl, dissociates cohesin from chromatin [31 – 33]. If the molecular function of cohesin involves topological embrace, the ring must be opened for both DNA entrapment and release.
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Nov 04, 2020 · Mis4Scc2 initially forms a tertiary complex with cohesin on DNA and promotes subsequent topological DNA entrapment by cohesin through its DNA binding activity. Furthermore, we show that mutations in the two DNA binding sites of Mis4 impair the chromosomal loading of cohesin.
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DNA binds to Ycs4 in the condensin bridged state. The hinge could additionally contribute to DNA binding. DNA binding promotes closure of the ATPase heads, resulting in DNA trapped within both the K and the S compartment. Proximity of ATPase heads promote ATP binding, release of Ycs4 and binding of Ycg1 to the Smc2 ATPase heads.
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Dec 01, 2013 · Biochemical reconstitution of topological DNA binding by the cohesin ring DNA using purified fission yeast cohesin and its loader complex, Mis4(Scc2)-Ssl3(Scc4) (Schizosaccharomyces pombe …
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Jun 19, 2017 · Cohesin is a conserved, ring-shaped protein complex that encircles sister chromatids and ensures correct chromosome segregation during mitosis and meiosis. It also plays a crucial role in the regulation of gene expression, DNA condensation, and DNA repair through both non-homologous end joining and homologous recombination. Cohesins are spatiotemporally regulated by the Scc2–Scc4 …
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Cohesin, a critical mediator of genome organization including sister chromatid cohesion, is a ring-shaped multi-subunit ATPase that topologically embraces DNA. Its loading and fun
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May 19, 2009 · Little is known about the Mis4/Ssl3 cohesin loader (orthologs of budding yeast Scc2/Scc4), a protein complex that is required for cohesin's association with chromosomes [3, 22, 23]. How the cohesin loader recognizes its binding sites on chromosomes, and how it promotes cohesin loading at these sites, are poorly understood.
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The resulting atomic model covered the two Psm head domains and adjacent coiled coil, bound to the kleisin N- and C-termini, as well as the Mis4 loader and 32bp of DNA. The structure gave unprecedented insight into mechanism. Mis4 was clamping the DNA onto the ATPase head domains, making widespread contacts with both Psm1 and Psm3.
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Sep 25, 2018 · (A) Effect of ADP/AlFx on loader binding to cohesin using DNA-bead assay. WT cohesin and an equimolar amount of loader was assayed for binding to DNA beads in the presence or ATP or ADP/AlFx. Silver-stained gel to assess cohesin (Psm3, Psm1, Rad21/Psc3) and loader (Mis4, Ssl3) bound to …
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Jun 19, 2017 · Cohesin is a conserved, ring-shaped protein complex that encircles sister chromatids and ensures correct chromosome segregation during mitosis and meiosis. It also plays a crucial role in the regulation of gene expression, DNA condensation, and DNA repair through both non-homologous end joining and homologous recombination. Cohesins are spatiotemporally regulated by the Scc2–Scc4 …
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