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Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes
Published on: December 20, 2014
Centriole reduplication during prolonged interphase requires procentriole maturation governed by Plk1
Jadranka Loncarek1, Polla Hergert, Alexey Khodjakov
1Division of Translational Medicine, Wadsworth Center, New York State Department of Health, Albany, NY 12201-0509, USA. loncarek@wadsworth.org
Current Biology : CB
|July 27, 2010
Summary
Centriole reduplication, leading to abnormal cell division and cancer, requires Polo-like kinase 1 (Plk1) activity. Plk1 activation during cell cycle arrest triggers centriole maturation and duplication, coordinating the centriole cycle with cell cycle progression.
Area of Science:
- Cell Biology
- Molecular Biology
- Cancer Research
Background:
- Supernumerary centrioles disrupt normal mitosis, promoting tumorigenesis.
- Centriole duplication must be tightly regulated and synchronized with the cell cycle.
- Centriole reduplication during prolonged interphase in transformed cells is poorly understood.
Purpose of the Study:
- To investigate the mechanisms underlying centriole reduplication during cell cycle arrest.
- To determine the role of Polo-like kinase 1 (Plk1) in centriole reduplication.
- To elucidate how Plk1 coordinates the centriole duplication cycle with the cell cycle.
Main Methods:
- Analysis of centriole reduplication in human cancerous and non-transformed cells during G2 and S phase arrests.
- Investigation of the role of activated Polo-like kinase 1 (Plk1) at the centrosome.
- Assessment of procentriole maturation and disengagement in the presence and absence of Plk1 activity.
Main Results:
- Centrioles consistently reduplicate in both cancerous and non-transformed human cells during G2 arrest.
- Plk1 activity is essential for centriole reduplication during G2 arrest.
- Activated Plk1 (phosphorylated at Thr210) at the centrosome is crucial for centriole reduplication during S arrest, promoting procentriole maturation and disengagement.
Conclusions:
- Polo-like kinase 1 (Plk1) plays a critical role in coordinating the centriole duplication cycle with the cell cycle.
- Plk1 activation during late S/G2 phases induces procentriole maturation, enabling completion of the centriole cycle independently of cell cycle progression.
- Understanding Plk1's role in centriole reduplication offers insights into preventing abnormal mitosis and tumorigenesis.
Related Concept Videos
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...
01:13Centrioles 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,"...
02:50Spindle 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...
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...
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...
02:57Attachment 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...