Cohesin loading and sliding | Journal of Cell Science
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 complex with chromosomes displays similarities with cohesin in its preference for convergent intergenes, but also follows additional determinants of binding that
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Figures and data in A Brownian ratchet model for DNA loop
Jul 26, 2021 · ATPase head disengagement and return of Scc2 Mis4 from the gripping state conformation to its extended crystal structure form results in loss of DNA interactions, turning the module into its 'slipping state'. (C) The Scc3-hinge module. The DNA binding surface of this module remains unaltered by ATP hydrolysis.
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In vitro loading of human cohesin on DNA by the human Scc2
Jun 12, 2012 · Isolation of h-Scc2/4 and h-Scc2/4 Derivatives. Using the baculovirus–Sf9 system, we cloned a number of h-Scc2 derivatives containing amino acids 1–500, 1–1,500, and 1–500 plus 1,542–2,375 (N + HEAT), all associated with h-Scc4 (Fig. 1 A and B).The sedimentation of these complexes through glycerol gradients, followed by SDS/PAGE analysis, revealed that they …
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Condensin complexes: understanding loop extrusion one
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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Jan 16, 2014 · In the absence of protein, or in the presence of Mis4 Scc2 /Ssl3 Scc4 only, no DNA was recovered (Fig. 2d). About 5% of input DNA was recovered when we performed the reaction in the presence of cohesin. The amount of bound DNA increased 5-fold when also Mis4 Scc2 /Ssl3 Scc4 was included. This suggests that the cohesin loader promotes salt-resistant cohesin …
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Cohesin dynamic association to chromatin and interfacing
Mar 16, 2018 · In vitro, second DNA capture depends on the Mis4/Ssl3 (Scc2/Scc4) complex, so far the only known factor able to recruit cohesin to chromatin. It is, therefore, reasonable to think that this complex may associate to replication sites to mediate cohesin "recycling" and entrap nascent chromatid in two consecutive steps.
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A Brownian ratchet model for DNA loop extrusion by the
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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DNA Binding by the Mis4 Scc2 Loader Promotes Topological
DNA Binding by the Mis4 Scc2 Loader Promotes Topological DNA Entrapment by the Cohesin Ring. 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 function on chromosomes require the Scc2-Scc4 loader.
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In vitro loading of human cohesin on DNA by the human Scc2
also loaded onto DNA at any stage of the cell cycle. Double stranded (ds)DNA breaks induce a damage response that increases the genomic binding of cohesin (8), whereas in yeast, cohesin is recruited to the damaged site (9). The role of cohesin following DNA damage occurs only after acetylation of Sccl (in yeast) or Smc3 (in vertebrates) (10).
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DNA Binding by the Mis4Scc2 Loader Promotes …
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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The CDK Pef1 and protein phosphatase 4 oppose each other
Jan 02, 2020 · Mis4 alone has DNA-binding activity and the C terminal hook domain is sufficient for stimulating DNA capture by cohesin in vitro (Chao et al., 2015; Murayama and Uhlmann, 2014). Mis4 makes multiple contacts with cohesin which may help cohesin conformational changes required for DNA capture ( Kikuchi et al., 2016 ; Murayama and Uhlmann, 2014 ).
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MCM2–7-dependent cohesin loading during S phase promotes
SCC2/4 then promotes the capture of a second DNA molecule by the fork-bound cohesin, thus establishing sister-chromatid cohesion (Figure 7). Components required for Okazaki fragment maturation and ligation and histone deposition are dispensable for cohesion establishment, suggesting that cohesion establishment can occur prior to these events.
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DNA Binding by the Mis4Scc2 Loader Promotes Topological
Nov 12, 2020 · Dataset DNA Binding by the Mis4Scc2 Loader Promotes Topological DNA Entrapment by
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DNA entry, exit and second DNA capture by cohesin
DNA binding of the BsSMC dimer. Nonetheless, a Loader Pds5-Wapl ATP hydrolysis DNA synthesis Initial DNA loading Second DNA capture Smc3 Smc1 Scc1 Scc3 KK Cohesin Pds5 Wapl Scc2 Scc4 dsDNA-dsDNA tethering Loader DNA sensor ATP ssDNA dsDNA Figure 1. A model for DNA entry, exit and second DNA capture by cohesin. To highlight the actions of
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DNA Binding by the Mis4 Scc2 Loader Promotes …
Nov 12, 2020 · DNA Binding by the Mis4Scc2 Loader Promotes Topological DNA Entrapment by the Cohesin Ring. Kurokawa and Murayama: Creator: MURAYAMA, Y (via Mendeley Data) …
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MCM2–7-dependent cohesin loading during S phase promotes
SCC2/4 then promotes the capture of a second DNA molecule by the fork-bound cohesin, thus establishing sister-chromatid cohesion (Figure 7). Components required for Okazaki fragment maturation and ligation and histone deposition are dispensable for cohesion establishment, suggesting that cohesion establishment can occur prior to these events.
Get a quoteBiochemical reconstitution of topological DNA binding by
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 …
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Topological in vitro loading of the budding yeast cohesin
Jul 26, 2021 · Next, the DNA engages Scc2 Mis4, which turns into its gripping state conformation as its own positive charges embrace the DNA. Together, the DNA and cohesin loader also establish contact with and close the kleisin N-gate. The DNA-induced Scc2 Mis4 conformational change creates a docking interface for Scc3 Psc3. The latter subunit joins the complex by …
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Biochemical reconstitution of topological DNA binding by
Dec 01, 2013 · c, DNA binding of Mis4 Scc2 –Ssl3 Scc4, using an electrophoretic mobility shift assay. d, Quantification of DNA binding by Mis4 Scc2 –Ssl3 Scc4 (left) and Mis4 Scc2 (right). Mean and standard
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Biochemical reconstitution of topological DNA binding by
Dec 01, 2013 · The cohesin loader binds to DNA. We purified the fission yeast Mis4 Scc2 /Ssl3 Scc4 cohesin loader complex after overexpression of its two subunits in fission yeast (Fig. 1a and Extended Data Fig. 1a).We used a similar strategy to purify the large Mis4 Scc2 subunit by itself. Based on its hydrodynamic properties, Mis4 Scc2 /Ssl3 Scc4 is a moderately elongated, …
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