ArticleNature protocols2012
A cell-free system for functional centromere and kinetochore assembly.
Article in Nature protocols, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
What it found
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The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
31 citing papers in PubMed, 52 citations in OpenAlex.
- Determinants of centromeric nucleosome conformation.bioRxiv : the preprint server for biology · 2026Article
- Phosphorylation of Xenopus M18BP1 governs centromeric localization and CENP-A nucleosome assembly.EMBO reports · 2026Article
- Reconstitution of genome remodeling usingFrontiers in cell and developmental biology · 2026Review
- Preserving centromere identity: right amounts of CENP-A at the right place and time.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2025Review
- Article
- Interactions with multiple inner kinetochore proteins determine mitotic localization of FACT.The Journal of cell biology · 2025Article
- Repression of CENP-A assembly in metaphase requires HJURP phosphorylation and inhibition by M18BP1.The Journal of cell biology · 2023Article
- Aberrant phase separation is a common killing strategy of positively charged peptides in biology and human disease.bioRxiv : the preprint server for biology · 2023Article
- Differential nuclear import sets the timing of protein access to the embryonic genome.Nature communications · 2022Article
- Structure of the human inner kinetochore CCAN complex and its significance for human centromere organization.Molecular cell · 2022Article
- Auto-inhibition of Mif2/CENP-C ensures centromere-dependent kinetochore assembly in budding yeast.The EMBO journal · 2020Article
- Cryoelectron Microscopy Structure of a Yeast Centromeric Nucleosome at 2.7 Å Resolution.Structure (London, England : 1993) · 2020Article
- Article
- CDK phosphorylation ofThe EMBO journal · 2019Article
- Measurement of Mesoscale Conformational Dynamics of Freely Diffusing Molecules with Tracking FCS.Biophysical journal · 2018Article
- Constitutive centromere-associated network contacts confer differential stability on CENP-A nucleosomes in vitro and in the cell.Molecular biology of the cell · 2018Article
- Nanoscale dynamics of centromere nucleosomes and the critical roles of CENP-A.Nucleic acids research · 2018Article
- Direct AFM Visualization of the Nanoscale Dynamics of Biomolecular Complexes.Journal of physics D: Applied physics · 2018Article
- RCC1 regulates inner centromeric composition in a Ran-independent fashion.Cell cycle (Georgetown, Tex.) · 2018Article
- Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
This protocol describes a cell-free system for studying vertebrate centromere and kinetochore formation. We reconstitute tandem arrays of centromere protein A (CENP-A) nucleosomes as a substrate for centromere and kinetochore assembly. These chromatin substrates are immobilized on magnetic beads and then incubated in Xenopus egg extracts that provide a source for centromere and kinetochore proteins and that can be cycled between mitotic and interphase cell cycle states. This cell-free system lends itself to use in protein immunodepletion, complementation and drug inhibition as a tool to perturb centromere and kinetochore assembly, cytoskeletal dynamics, DNA modification and protein post-translational modification. This system provides a distinct advantage over cell-based investigations in which perturbing centromere and kinetochore function often results in lethality. After incubation in egg extract, reconstituted CENP-A chromatin specifically assembles centromere and kinetochore proteins, which locally stabilize microtubules and, on microtubule depolymerization with nocodazole, activate the mitotic checkpoint. A typical experiment takes 3 d.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.