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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.
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M-Cdk Drives Transition Into Mitosis

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Related Experiment Video

Updated: May 7, 2026

Combining Mitotic Cell Synchronization and High Resolution Confocal Microscopy to Study the Role of Multifunctional Cell Cycle Proteins During Mitosis

Combining Mitotic Cell Synchronization and High Resolution Confocal Microscopy to Study the Role of Multifunctional Cell Cycle Proteins During Mitosis

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NuMA localization, stability, and function in spindle orientation involve 4.1 and Cdk1 interactions.

Lindsey Seldin¹, Nicholas D Poulson, Henry P Foote

¹Department of Cell Biology, Duke University Medical Center, Durham, NC 27710 Department of Dermatology, Duke University Medical Center, Durham, NC 27710.

Molecular Biology of the Cell

|October 11, 2013

Summary

This summary is machine-generated.

Nuclear mitotic apparatus (NuMA) protein is crucial for spindle orientation in keratinocytes. NuMA recruits dynactin and stabilizes at the cell cortex via its 4.1-binding domain, ensuring proper cell division.

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Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

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Published on: March 3, 2016

Area of Science:

Cell Biology
Epithelial Biology
Cytoskeletal Dynamics

Background:

Stratification of the epidermis relies on asymmetric cell divisions.
A protein complex involving LGN, NuMA, and dynein/dynactin is essential for spindle orientation.
Mechanisms governing the cortical recruitment of these proteins are not fully understood.

Purpose of the Study:

To elucidate the requirements for cortical recruitment of the NuMA-LGN-dynein/dynactin complex in keratinocytes.
To investigate the role of NuMA's 4.1-binding domain in its cortical localization and function.
To understand the regulation of NuMA cortical localization during mitosis.

Main Methods:

Immunofluorescence microscopy to visualize protein localization.
Fluorescence recovery after photobleaching (FRAP) to assess protein dynamics.
Functional assays to evaluate spindle orientation defects.

Main Results:

NuMA is required for dynactin recruitment to the keratinocyte cell cortex.
NuMA's cortical localization depends on LGN, but LGN interaction alone is insufficient.
The 4.1-binding domain of NuMA is critical for stabilizing its cortical interaction, and its loss causes spindle orientation defects.
Cortical NuMA levels increase during anaphase, regulated by Cdk1 activity and specific NuMA residues, independently of LGN and 4.1 interactions.

Conclusions:

NuMA plays a dual role in cortical recruitment: LGN-dependent localization and 4.1-mediated stabilization.
Distinct mechanisms govern NuMA cortical localization at different mitotic stages.
NuMA cortical stability is vital for generating forces necessary for spindle orientation during epidermal stratification.