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Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins
Published on: March 3, 2016
beta-Catenin is a Nek2 substrate involved in centrosome separation
Shirin Bahmanyar1, Daniel D Kaplan, Jennifer G Deluca
1Department of Biological Sciences, Stanford University, Stanford, California 94305, USA.
Genes & Development
|December 19, 2007
Summary
Beta-catenin regulates centrosome separation by binding to the intercentrosomal linker. Its stabilization, seen in cancers, triggers centrosome splitting, revealing a new role in cell division.
Area of Science:
- Cell Biology
- Molecular Biology
- Cancer Research
Background:
- Beta-catenin is crucial for cell adhesion and gene transcription.
- Beta-catenin is essential for establishing a bipolar mitotic spindle.
- Centrosome separation is regulated by Nek2 kinase, C-Nap1, and Rootletin.
Purpose of the Study:
- To investigate the role of beta-catenin in centrosome function and separation.
- To determine how beta-catenin interacts with the centrosome linker complex.
- To understand the implications of beta-catenin stabilization in cancer for centrosome dynamics.
Main Methods:
- Immunofluorescence microscopy to visualize protein localization.
- Co-immunoprecipitation assays to study protein interactions.
- Analysis of protein phosphorylation by Nek2 kinase.
- Experimental manipulation of beta-catenin stability and Nek2 activity.
Main Results:
- Beta-catenin is a component of interphase centrosomes, localizing with Rootletin.
- Stabilization of beta-catenin induces centrosome splitting, mimicking cancer mutations.
- Beta-catenin binds to and is phosphorylated by Nek2 kinase.
- During mitosis, beta-catenin relocates to spindle poles independently of Rootletin, facilitating centrosome separation.
Conclusions:
- Beta-catenin is a component of the intercentrosomal linker.
- Beta-catenin plays a novel role in regulating mitotic centrosome separation.
- Aberrant beta-catenin function contributes to centrosome abnormalities in cancer.
Related Concept Videos
01:23Catenins
Catenins are characterized by multiple binding domains and dynamic structures that allow them to function as linker proteins in cell junction complexes. All catenins, except α-catenin, contain a characteristic protein sequence called the armadillo repeat and are therefore also called armadillo proteins.
Catenins in Cell Junctions
Catenins bind to cell adhesion molecules such as cadherins and link them to different cytoskeletal proteins depending on the type of cell junction. At the adherens...
Catenins in Cell Junctions
Catenins bind to cell adhesion molecules such as cadherins and link them to different cytoskeletal proteins depending on the type of cell junction. At the adherens...
01:13Centrioles and Centrosomes
Most animal cells comprise a pair of centrioles together called a centrosome. The cell duplicates its centrosome and contains two centrosomes side-by-side, which begin to move apart during the prophase. As the centrosomes migrate to two different sides of the cell, microtubules start extending from each centrosome toward the other end. The mitotic spindle is composed of the centrosomes and their emerging microtubules.
Near the end of the prophase, also called late prophase or "prometaphase,"...
Near the end of the prophase, also called late prophase or "prometaphase,"...
02:25Centrosome Duplication
The primary microtubule organizing center (MTOC) in animal cells is the centrosome. A centrosome has two cylindrical centrioles at its core. Each centriole consists of nine sets of three microtubules held together by proteins. The centrioles are positioned at right angles to each other and surrounded by a shapeless protein cloud called the pericentriolar matrix, or pericentriolar material (PCM).
To ensure that each daughter cell receives a centrosome after cell division, centrosome duplication...
To ensure that each daughter cell receives a centrosome after cell division, centrosome duplication...
02:25Centrosome Duplication
The primary microtubule organizing center (MTOC) in animal cells is the centrosome. A centrosome has two cylindrical centrioles at its core. Each centriole consists of nine sets of three microtubules held together by proteins. The centrioles are positioned at right angles to each other and surrounded by a shapeless protein cloud called the pericentriolar matrix, or pericentriolar material (PCM).
To ensure that each daughter cell receives a centrosome after cell division, centrosome duplication...
To ensure that each daughter cell receives a centrosome after cell division, centrosome duplication...
01:26Tension Response at Adherens Junctions
The adherens junctions that anchor cells together are multi-protein complexes that dynamically adapt to mechanical stimuli such as tensile forces and shear stress. Mechanosensory proteins in these junctions can sense such mechanical stimuli and undergo a shift in their conformation, resulting in an altered function — a process called mechanotransduction.
α-Catenin as a Mechanosensory Protein
The α-catenin of adherens junctions is an allosteric protein with three VH (vinculin homology) domains...
α-Catenin as a Mechanosensory Protein
The α-catenin of adherens junctions is an allosteric protein with three VH (vinculin homology) domains...
02:17Separation of Sister Chromatids
At the transition from prophase to metaphase, there is a reduction in cohesion along the chromosomal arms, resulting in the resolution of sister chromatids. However, residual cohesin connections remain to hold the sister chromatids together until the transition from metaphase to anaphase. The residual connection prevents any premature separation of sister chromatids, blocking the risks of aneuploidy within the daughter cells.
At the onset of anaphase, separase, a proteolytic enzyme, is...
At the onset of anaphase, separase, a proteolytic enzyme, is...