Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Attachment of Sister Chromatids02:57

Attachment of Sister Chromatids

3.3K
As cells progress into mitosis, the nuclear envelope breaks down, and the condensed chromosomes are exposed to the array of bipolar microtubules of the mitotic spindle. The kinetochore, a large, disc-shaped protein complex, is present at the centromere region of the sister chromatids and acts as a binding site for the microtubules.  Usually, the plus-end of a single microtubule is embedded within the kinetochore. However, some kinetochores first establish lateral contact with the side-wall...
3.3K
Spindle Assembly02:50

Spindle Assembly

3.7K
Spindle assembly occurs through three, often coexisting, pathways – the centrosome-mediated pathway, the chromatin-mediated pathway, and the microtubule-mediated pathway – collectively contributing to form a robust spindle apparatus.
In most cells, centrosomes are the primary microtubule nucleation centers. In the centrosome-mediated pathway, the G2-prophase transition triggers centrosome maturation and increased microtubule nucleation. Progressive nucleation results in a...
3.7K
The Spindle Assembly Checkpoint02:19

The Spindle Assembly Checkpoint

3.2K
The spindle assembly checkpoint is a molecular surveillance mechanism ensuring the fidelity of chromosome segregation during anaphase. The checkpoint monitors the completion of all the prerequisite steps before chromosome segregation to determine whether the segregation process should proceed or be delayed.
Many proteins function together to control the spindle assembly checkpoint. Mutations affecting these proteins may allow cells to proceed into anaphase prematurely, resulting in the...
3.2K
Assembly of Cytoskeletal Filaments01:18

Assembly of Cytoskeletal Filaments

20.0K
Cytoskeletal filaments are polymeric forms of smaller protein subunits. However, individual cytoskeletal filaments may easily disassemble or associate with other similar filaments to form rigid structures. Microfilaments, made of actin monomers, rely on actin-binding proteins to form bundles and create networks of individual actin filaments. Microtubules rely on microtubule-associated proteins (MAPs) to form sturdy cylindrical structures. However, the proteins involved in forming complex...
20.0K
Protein Complex Assembly02:41

Protein Complex Assembly

10.6K
Proteins can form homomeric complexes with another unit of the same protein or heteromeric complexes with different types.  Most protein complexes self-assemble spontaneously via ordered pathways, while some proteins need assembly factors that guide their proper assembly. Despite the crowded intracellular environment, proteins usually interact with their correct partners and form functional complexes.
Many viruses self-assemble into a fully functional unit using the infected host cell to...
10.6K
Forces Acting on Chromosomes02:11

Forces Acting on Chromosomes

3.3K
During mitosis, chromosome movements occur through the interplay of multiple piconewton level forces. In prometaphase, these forces help in chromosome assembly or congression at the equatorial plane, eventually leading to their alignment at the metaphase plate. The forces acting on the chromosomes are space and time-dependent; therefore, they vary with the position of the chromosomes as the cell progresses through mitosis. 
Microtubules and motor proteins exert two types of forces on...
3.3K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Brieflow: an integrated computational pipeline for high-throughput analysis of optical pooled screening data.

Nature communications·2026
Same author

5' UTR length shapes alternative N-terminal protein isoforms across cancers and in rare disease.

EMBO reports·2026
Same author

The dynamics of centromere assembly and disassembly during quiescence.

The Journal of cell biology·2026
Same author

Global stabilization of the transcriptome in mitotic cells.

The EMBO journal·2026
Same author

5' UTR length regulates alternative N-terminal protein isoform production in health and disease.

bioRxiv : the preprint server for biology·2026
Same author

Regulation of mRNA decay and translation during the mammalian cell cycle.

RNA (New York, N.Y.)·2026

Related Experiment Video

Updated: Jul 6, 2025

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins
05:35

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

Published on: March 3, 2016

15.2K

Higher-order protein assembly controls kinetochore formation.

Gunter B Sissoko1,2, Ekaterina V Tarasovetc3, Océane Marescal1,2

  • 1Whitehead Institute for Biomedical Research, Cambridge, MA, USA.

Nature Cell Biology
|January 3, 2024
PubMed
Summary

Oligomerization of the inner kinetochore protein CENP-T, not individual molecules, drives outer kinetochore assembly. This density-dependent mechanism ensures precise chromosome segregation during mitosis.

More Related Videos

Protein Purification Technique that Allows Detection of Sumoylation and Ubiquitination of Budding Yeast Kinetochore Proteins Ndc10 and Ndc80
12:28

Protein Purification Technique that Allows Detection of Sumoylation and Ubiquitination of Budding Yeast Kinetochore Proteins Ndc10 and Ndc80

Published on: May 3, 2015

12.2K
Reconstitution of Basic Mitotic Spindles in Spherical Emulsion Droplets
10:52

Reconstitution of Basic Mitotic Spindles in Spherical Emulsion Droplets

Published on: August 13, 2016

9.8K

Related Experiment Videos

Last Updated: Jul 6, 2025

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins
05:35

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

Published on: March 3, 2016

15.2K
Protein Purification Technique that Allows Detection of Sumoylation and Ubiquitination of Budding Yeast Kinetochore Proteins Ndc10 and Ndc80
12:28

Protein Purification Technique that Allows Detection of Sumoylation and Ubiquitination of Budding Yeast Kinetochore Proteins Ndc10 and Ndc80

Published on: May 3, 2015

12.2K
Reconstitution of Basic Mitotic Spindles in Spherical Emulsion Droplets
10:52

Reconstitution of Basic Mitotic Spindles in Spherical Emulsion Droplets

Published on: August 13, 2016

9.8K

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Accurate chromosome segregation during mitosis relies on precise kinetochore-microtubule interactions.
  • Mechanisms preventing outer kinetochore formation at low CENP-A nucleosome density remain unclear.

Purpose of the Study:

  • Investigate the role of higher-order assembly of the inner kinetochore protein CENP-T in kinetochore formation.
  • Determine how CENP-T density impacts outer kinetochore recruitment.

Main Methods:

  • Generated oligomers of CENP-T using two distinct genetic engineering systems in human cells.
  • Assessed interactions between oligomerized CENP-T and outer kinetochore proteins in vitro and in vivo.
  • Quantified recruitment levels of outer kinetochore components to monomeric versus oligomeric CENP-T.

Main Results:

  • Oligomers of CENP-T, mimicking centromeric density, robustly formed functional kinetochore-like particles.
  • Oligomerized CENP-T recruited significantly higher levels of outer kinetochore components compared to monomeric CENP-T.
  • This density-dependent recruitment was observed both in cellular and in vitro systems.

Conclusions:

  • The density of CENP-T, rather than individual protein interactions, is critical for initiating outer kinetochore assembly.
  • This density dependence restricts outer kinetochore recruitment to centromeres where CENP-A density is high.
  • The findings elucidate a key mechanism for ensuring faithful chromosome segregation during vertebrate mitosis.