Structure of the Pds5-Scc1 Complex and Implications for Cohesin Function

Muir, Kyle; Kschonsak, Marc; Li, Yan; Metz, J; Haering, Christian H.; Panne, Daniel

doi:10.1016/j.celrep.2016.01.078

Cited by 49 publications

(46 citation statements)

References 56 publications

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“…Thus, the curvature of Scc2 is a defined, conserved feature of the cohesin loader. The highly bent structure of Scc2 is reminiscent of the structures of Scc3 [stromal antigen 1 or 2 (SA1/2) in vertebrates] and Pds5 (8,9,27,35,36) (Fig. 1 C and D).…”

Section: Resultsmentioning

confidence: 92%

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Crystal structure of the cohesin loader Scc2 and insight into cohesinopathy

Kikuchi

Borek

Otwinowski

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

117

125

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The ring-shaped cohesin complex topologically entraps chromosomes and regulates chromosome segregation, transcription, and DNA repair. The cohesin core consists of the structural maintenance of chromosomes 1 and 3 (Smc1-Smc3) heterodimeric ATPase, the kleisin subunit sister chromatid cohesion 1 (Scc1) that links the two ATPase heads, and the Scc1-bound adaptor protein Scc3. The sister chromatid cohesion 2 and 4 (Scc2-Scc4) complex loads cohesin onto chromosomes. Mutations of cohesin and its regulators, including Scc2, cause human developmental diseases termed cohesinopathy. Here, we report the crystal structure of Chaetomium thermophilum (Ct) Scc2 and examine its interaction with cohesin. Similar to Scc3 and another Scc1-interacting cohesin regulator, precocious dissociation of sisters 5 (Pds5), Scc2 consists mostly of helical repeats that fold into a hook-shaped structure. Scc2 binds to Scc1 through an N-terminal region of Scc1 that overlaps with its Pds5-binding region. Many cohesinopathy mutations target conserved residues in Scc2 and diminish Ct Scc2 binding to Ct Scc1. Pds5 binding to Scc1 weakens the Scc2-Scc1 interaction. Our study defines a functionally important interaction between the kleisin subunit of cohesin and the hook of Scc2. Through competing with Scc2 for Scc1 binding, Pds5 might contribute to the release of Scc2 from loaded cohesin, freeing Scc2 for additional rounds of loading.transcription | cohesinopathy | X-ray crystallography | cohesin loading | HEAT repeat C ohesin consists of four core subunits: structural maintenance of chromosomes 1 and 3 (Smc1 and Smc3), and sister chromatid cohesion 1 and 3 (Scc1 and Scc3). (1-5). Smc1 and Smc3 are related ATPases that heterodimerize through their hinge domains. The C-terminal winged helix domain and the N-terminal helical domain (NHD) of Scc1 bind to the Smc1 ATPase head and a coiled-coil segment adjacent to the Smc3 ATPase head, respectively, forming a tripartite ring (6, 7). Scc3 contains Huntingtin-elongation factor 3-protein phosphatase 2A-TOR1 (HEAT) repeats, binds to the middle region of Scc1, and provides a landing pad for several cohesin regulators (8, 9). Cohesin can topologically trap DNA inside its ring (10), thereby regulating many facets of chromosome biology. During interphase, cohesin mediates the formation of chromosome loops that impact transcription (11). During S phase, cohesin physically links replicated chromosomes to establish sister-chromatid cohesion (12, 13), which is a prerequisite for faithful chromosome segregation during mitosis. Finally, cohesin contributes to homologydirected repair of DNA breaks, in part, through holding the sister chromatid in proximity of the breaks and presenting it as the repair template (14).For cohesin to accomplish these diverse tasks, its association with chromosomes has to be tightly controlled both spatially and temporally. Cohesin dynamics on chromosomes are regulated by a set of accessory proteins, including the Scc2-Scc4 loading complex and the releasing complex comprising Pds5 and wing...

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Section: Resultsmentioning

confidence: 92%

“…Scc2 consists almost entirely of helical repeats that fold into a molecular hook. The overall structure of Scc2 is similar to two other Scc1-binding HEAT repeat proteins, Scc3 and Pds5, which also adopt highly bent structures (8,9,27,35,36). Like Scc3 and Pds5, Scc2 interacts with Scc1.…”

mentioning

confidence: 77%

Crystal structure of the cohesin loader Scc2 and insight into cohesinopathy

Kikuchi

Borek

Otwinowski

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

117

125

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“…However, these protocols led to the automatic solution of 20 unique structures in 2015. [47,56] While treating all user samples as a whole has led to important developments on the beamline, [40] it can mask the various trends that occur at the level of specific projects. The same analysis is therefore presented for three individual projects that represent different modes of using the beamline (Figure 7(b)-(d)).…”

Section: How Is the Beamline Used?mentioning

confidence: 99%

“…Of these processed data sets, two demonstrated a diffraction limit beyond 4 Å and one was used to solve the structure. [56] As these crystals grew in multiple plates, the X-ray centring routine allowed the best positions to be sampled consistently. Screening this number of crystals using manual centring on multiple positions is time consuming and samples fewer positions.…”

Section: How Is the Beamline Used?mentioning

confidence: 99%

Fully automatic macromolecular crystallography: the impact of MASSIF-1 on the optimum acquisition and quality of data

Bowler

Svensson

Nurizzo

2016

Crystallography Reviews

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Automation is beginning to transform the way data are collected in almost all scientific disciplines. The combination of robotics and software now allows data to be collected consistently and reproducibly, eliminating human error and boredom. This approach has been applied to macromolecular crystallography at MASSIF-1, a fully automated beamline at the European Synchrotron Radiation Facility (ESRF). Considerable human effort is still dedicated to evaluating protein crystals in order to find the few crystals that diffract well or collecting hundreds of data sets to screen potential new drug candidates. The combination of ESRF-developed robotic sample handling and advanced software protocols now provides a new tool to structural biologists. Not only is the beamline used efficiently, running 24 h a day without getting tired, data collection is also performed consistently by an expert system, often better than with a human operator. In this review, we will focus on the impact this level of automation has had on the optimum acquisition of data from crystals of biological macromolecules.

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“…This opened full automation to any sample presented in any mount and has provided a new tool to structural biologists, allowing the process of collecting hundreds of data sets or screening hundreds of crystals to be 'outsourced', freeing their time and often collecting better data . At the time of writing, the beamline has processed more than 39,000 samples representing a wide range of projects, from those that require extensive screening to find the best diffracting crystal (Na et al, 2017;Sorigué et al, 2017;Naschberger et al 2017) to small molecule fragment screening (Cheeseman et al, 2017;Hiruma et al, 2017) and experimental phasing at high and low resolutions (Kharde et al, 2015;Muir et al, 2016). The beamline is able to deal with a wide range of samples by combining parameters provided by the user with information gathered during processing.…”

Section: Introductionmentioning

confidence: 99%

Recent improvements to the automatic characterization and data collection algorithms on MASSIF-1

Svensson

Gilski

Nurizzo

et al. 2017

Preprint

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SynopsisSignificant improvements in the sample location, characterisation and data collection algorithms on the autonomous ESRF beamline MASSIF-1 are described. The workflows now include dynamic beam diameter adjustment and multi-position and multi-crystal data collections.. CC-BY-ND 4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/236596 doi: bioRxiv preprint first posted online 2 Abstract Macromolecular crystallography (MX) is now a mature and widely used technique essential in the understanding of biology and medicine. Increases in computing power combined with robotics have enabled not only large numbers of samples to be screened and characterised but also for better decisions to be taken on data collection itself. This led to the development of MASSIF-1 at the ESRF, the world's first beamline to run fully automatically while making intelligent decisions taking user requirements into account. Since opening in late 2014 the beamline has now processed over 39,000 samples. Improvements have been made to the speed of the sample handling robotics and error management within the software routines.The workflows initially put in place, while highly innovative at the time, have been expanded to include increased complexity and additional intelligence using the information gathered during characterisation, this includes adapting the beam diameter dynamically to match the diffraction volume within the crystal. Complex multi-position and multi-crystal data collections are now also integrated into the selection of experiments available. This has led to increased data quality and throughput allowing even the most challenging samples to be treated automatically.Key words: X-ray centring; synchrotron instrumentation; macromolecular crystallography; automation; helical data collection; multiple crystal data collection . CC-BY-ND 4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/236596 doi: bioRxiv preprint first posted online 3 IntroductionAutomation is transforming the way scientific data are collected, allowing large amounts of high quality data to be gathered in a consistent manner (Quintana & Plätzer, 2015;Foster, 2005). Advances in robotics and software have been key in these developments and have had a particular impact on structural biology, allowing multiple constructs to be screened and purified (Camper & Viola, 2009;Hart & Waldo, 2013;Vijayachandran et al., 2011); huge numbers of crystallisation experiments to be performed (Elsliger et al., 2010;Ferrer et al., 2013;Heinemann et al., 2003;Joachimiak, 2009;Calero et al., 2014), samples to be mounted at synchrotrons (Cipriani et al., 2006; Cohen et al., 2002;Jacquamet et al., 2009;Papp et al., 2017;Snell et al., 2004), data to be analysed and processed (Bourenkov & Popov, 2010;Holton & Alber, 2004;Incardona et al., ...

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Structure of the Pds5-Scc1 Complex and Implications for Cohesin Function

Cited by 49 publications

References 56 publications

Crystal structure of the cohesin loader Scc2 and insight into cohesinopathy

Crystal structure of the cohesin loader Scc2 and insight into cohesinopathy

Fully automatic macromolecular crystallography: the impact of MASSIF-1 on the optimum acquisition and quality of data

Recent improvements to the automatic characterization and data collection algorithms on MASSIF-1

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