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Attachment of Sister Chromatids

Attachment of Sister Chromatids

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 of a...

Spindle Assembly

Spindle Assembly

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 microtubule array...

Forces Acting on Chromosomes

Forces Acting on Chromosomes

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...

Anaphase A and B

Anaphase A and B

Microtubules form through the end-to-end polymerization of tubulin heterodimers. Kinetochore microtubules originate from the spindle poles, and their plus-ends connect with the kinetochores on sister-chromatids. Ndc80 protein complexes, present on the kinetochore, form low-affinity links with the plus end of these kinetochore microtubules.
Plus-end depolymerization releases tubulin heterodimers from the terminal region of the microtubule. As tubulin subunits are lost, the Ndc80 complexes detach...

The Spindle Assembly Checkpoint

The Spindle Assembly Checkpoint

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...

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Related Experiment Video

Updated: Jul 13, 2026

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Published on: October 27, 2011

The DT40 system as a tool for analyzing kinetochore assembly.

Masahiro Okada¹, Tetsuya Hori, Tatsuo Fukagawa

¹Department of Molecular Genetics, National Institute of Genetics and The Graduate University for Advanced Studies, Mishima, Shizuoka 411-8540, Japan.

Sub-Cellular Biochemistry

|July 13, 2007

Summary

Kinetochore assembly in vertebrates is complex. This review highlights recent advances in understanding kinetochore mechanisms using the DT40 cell system, a powerful model for chromosome segregation research.

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Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

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Last Updated: Jul 13, 2026

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Published on: October 27, 2011

Studying Mitotic Checkpoint by Illustrating Dynamic Kinetochore Protein Behavior and Chromosome Motion in Living Drosophila Syncytial Embryos

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Published on: June 14, 2012

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

Published on: March 9, 2022

Area of Science:

Cell Biology
Molecular Biology
Genetics

Background:

The kinetochore is a protein complex essential for chromosome segregation, mediating spindle microtubule attachment.
While budding yeast is a model, vertebrate kinetochore components are difficult to identify via homology, necessitating vertebrate system studies.
Understanding kinetochore assembly is crucial for comprehending eukaryotic chromosome segregation.

Purpose of the Study:

To review recent advances in understanding kinetochore assembly mechanisms.
To highlight the utility of the DT40 cell system for studying kinetochore assembly.
To discuss the challenges and strategies in identifying vertebrate kinetochore components.

Main Methods:

Review of experimental strategies including RNA interference (RNAi), knockout mice, Drosophila genetics, C. elegans studies, and Xenopus egg extracts.
Focus on the application of the DT40 cell system for kinetochore component identification and characterization.
Analysis of recent findings from the authors' laboratory.

Main Results:

The DT40 cell system provides a powerful and reliable platform for investigating kinetochore assembly.
Advances have been made in identifying and characterizing kinetochore components in vertebrate systems.
Comparative studies reveal complexities in kinetochore assembly across eukaryotes.

Conclusions:

The DT40 system is instrumental in dissecting vertebrate kinetochore assembly mechanisms.
Continued research in vertebrate models is essential for a complete understanding of chromosome segregation.
Identifying kinetochore components is key to understanding faithful chromosome inheritance.