The spindle checkpoint, also known as the metaphase-to-anaphase transition, the spindle assembly checkpoint (SAC), the metaphase checkpoint, or the mitotic checkpoint, is a cell cycle checkpoint during metaphase of mitosis or meiosis that prevents the separation of the duplicated chromosomes (anaphase) until each chromosome is properly attached to the spindle. To achieve proper segregation, the two kinetochores on the sister chromatids must be attached to opposite spindle poles (bipolar orientation). Only this pattern of attachment will ensure that each daughter cell receives one copy of the chromosome. The defining biochemical feature of this checkpoint is the stimulation of the anaphase-promoting complex by M-phase cyclin-CDK complexes, which in turn causes the proteolytic destruction of cyclins and proteins that hold the sister chromatids together.
Overview and importance The beginning of metaphase is characterized by the connection of the microtubules to the kinetochores of the chromosomes, as well as the alignment of the chromosomes in the middle of the cell. Each chromatid has its own kinetochore, and all of the microtubules that are bound to kinetochores of sister chromatids radiate from opposite poles of the cell. These microtubules exert a pulling force on the chromosomes towards the opposite ends of the cells, while the cohesion between the sister chromatids opposes this force. At the metaphase to anaphase transition, this cohesion between sister chromatids is dissolved, and the separated chromatids are pulled to opposite sides of the cell by the spindle microtubules. The chromatids are further separated by the physical movement of the spindle poles themselves. Premature dissociation of the chromatids can lead to chromosome missegregation and aneuploidy in the daughter cells. Thus, the job of the spindle checkpoint is to prevent this transition into anaphase until the chromosomes are properly attached, before the sister chromatids separate. In order to preserve the cell's identity and proper function, it is necessary to maintain the appropriate number of chromosomes after each cell division. An error in generating daughter cells with fewer or greater number of chromosomes than expected (a situation termed aneuploidy), may lead in best case to cell death, or alternatively it may generate catastrophic phenotypic results. Examples include:
In cancer cells, aneuploidy is a frequent event, indicating that these cells present a defect in the machinery involved in chromosome segregation, as well as in the mechanism ensuring that segregation is correctly performed. In humans, Down syndrome appears in children carrying in their cells one extra copy of chromosome 21, as a result of a defect in chromosome segregation during meiosis in one of the progenitors. This defect will generate a gamete (spermatozoide or oocyte) with an extra chromosome 21. After fertilisation, this gamete will generate an embryo with three copies of chromosome 21.
Discovery of the spindle assembly checkpoint (SAC)
Zirkle (in 1970) was one of the first researchers to observe that, when just one chromosome is retarded to arrive at the metaphase plate, anaphase onset is postponed until some minutes after its arrival. This observation, together with similar ones, suggested that a control mechanism exists at the metaphase-to-anaphase transition. Using drugs such as nocodazole and colchicine, the mitotic spindle disassembles and the cell cycle is blocked at the metaphase-to-anaphase transition. Using these drugs (see the review from Rieder and Palazzo in 1992), the putative control mechanism was named Spindle Assembly Checkpoint (SAC). This regulatory mechanism has been intensively studied since. Using different types of genetic studies, it has been established that diverse kinds of defects are able to activate the SAC: spindle depolymerization, the presence of dicentric chromosomes (with two centromeres), centromeres segregating in an aberrant way, defects in the spindle pole bodies in S. cerevisiae, defects in the kinetochore proteins, mutations in the centromeric DNA or defects in the molecular motors active during mitosis. A summary of these observations can be found in the article from Hardwick and collaborators in 1999. Using its own observations, Zirkle was the first to propose that "some (…) substance, necessary for the cell to proceed to anaphase, appears some minutes after C (moment of the arrival of the last chromosome to the metaphase plate), or after a drastic change in the cytoplasmic condition, just at C or immediately after C", suggesting that this function is located on kinetochores unattached to the mitotic spindle. McIntosh extended this proposal, suggesting that one enzyme sensitive to tension located at the centromeres produces an inhibitor to the anaphase onset when the two sister kinetochores are not under bipolar tension. Indeed, the available data suggested that the signal "wait to enter in anaphase" is produced mostly on or close to unattached kinetochores. However, the primary event associated to the kinetochore attachment to the spindle, which is able to inactivate the inhibitory signal and release the metaphase arrest, could be either the acquisition of microtubules by the kinetochore (as proposed by Rieder and collaborators in 1995), or the tension stabilizing the anchoring of microtubules to the kinetochores (as suggested by the experiments realized at Nicklas' lab). Subsequent studies in cells containing two independent mitotic spindles in a sole cytoplasm showed that the inhibitor of the metaphase-to-anaphase transition is generated by unattached kinetochores and is not freely diffusible in the cytoplasm. Yet in the same study it was shown that, once the transition from metaphase to anaphase is initiated in one part of the cell, this information is extended all along the cytoplasm, and can overcome the signal "wait to enter in anaphase" associated to a second spindle containing unattached kinetochores.
Background on sister chromatid duplication, cohesion, and segregation
Cell division: duplication of material and distribution to daughter cells
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