Cell Cycle

A cell cycle is a series of events that takes place in a cell as it grows and divides. A cell spends most of its
time in what is called interphase, and during this time it grows, replicates its chromosomes, and
prepares for cell division. The cell then leaves interphase, undergoes mitosis, and completes its division.
The resulting cells, known as daughter cells, each enter their own interphase and begin a new round of
the cell cycle.

The cell cycle, or cell-division cycle, is the series of events that take place in a cell that cause it to divide
into two daughter cells. These events include the duplication of its DNA (DNA replication) and some of
its organelles, and subsequently the partitioning of its cytoplasm and other components into two
daughter cells in a process called cell division.

In cells with nuclei (eukaryotes), (i.e., animal, plant, fungal, and protist cells), the cell cycle is divided into
two main stages: interphase and the mitotic (M) phase (including mitosis and cytokinesis). During
interphase, the cell grows, accumulating nutrients needed for mitosis, and replicates its DNA and some
of its organelles. During the mitotic phase, the replicated chromosomes, organelles, and cytoplasm
separate into two new daughter cells. To ensure the proper replication of cellular components and
division, there are control mechanisms known as cell cycle checkpoints after each of the key steps of the
cycle that determine if the cell can progress to the next phase.

In cells without nuclei (prokaryotes), (i.e., bacteria and archaea), the cell cycle is divided into the B, C,
and D periods. The B period extends from the end of cell division to the beginning of DNA replication.
DNA replication occurs during the C period. The D period refers to the stage between the end of DNA
replication and the splitting of the bacterial cell into two daughter cells.[1]

The cell-division cycle is a vital process by which a single-celled fertilized egg develops into a mature
organism, as well as the process by which hair, skin, blood cells, and some internal organs are renewed.
After cell division, each of the daughter cells begin the interphase of a new cycle. Although the various
stages of interphase are not usually morphologically distinguishable, each phase of the cell cycle has a
distinct set of specialized biochemical processes that prepare the cell for initiation of the cell division.

hases[edit]

The eukaryotic cell cycle consists of four distinct phases: G1 phase, S phase (synthesis), G2 phase
(collectively known as interphase) and M phase (mitosis and cytokinesis). M phase is itself composed of
two tightly coupled processes: mitosis, in which the cell's nucleus divides, and cytokinesis, in which the
cell's cytoplasm divides forming two daughter cells. Activation of each phase is dependent on the proper
progression and completion of the previous one. Cells that have temporarily or reversibly stopped
dividing are said to have entered a state of quiescence called G0 phase.

Schematic of the cell cycle. Outer ring: I = Interphase, M = Mitosis; inner ring: M = Mitosis, G1 = Gap 1,
G2 = Gap 2, S = Synthesis; not in ring: G0 = Gap 0/Resting[2]

State Phase Abbreviation Description

Resting Gap 0 G0 A phase where the cell has left the cycle and has stopped dividing.

Interphase Gap 1 G1 Cells increase in size in Gap 1. The G1 checkpoint control mechanism
ensures that everything is ready for DNA synthesis.

Synthesis S DNA replication occurs during this phase.

Gap 2 G2 During the gap between DNA synthesis and mitosis, the cell will continue to grow. The
G2 checkpoint control mechanism ensures that everything is ready to enter the M (mitosis) phase and
divide.

Cell division Mitosis M Cell growth stops at this stage and cellular energy is focused on the
orderly division into two daughter cells. A checkpoint in the middle of mitosis (Metaphase Checkpoint)
ensures that the cell is ready to complete cell division.

After cell division, each of the daughter cells begin the interphase of a new cycle. Although the various
stages of interphase are not usually morphologically distinguishable, each phase of the cell cycle has a
distinct set of specialized biochemical processes that prepare the cell for initiation of cell division.

Cell cycle checkpoint

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Steps of the cell cycle. The restriction point occurs between the G1 and S phases of interphase. The G2-
M checkpoint occurs between the G2 and M phases. The spindle checkpoint occurs during the M phase.
Key cyclins associated with each phase are shown.

Cell cycle checkpoints are control mechanisms in the eukaryotic cell cycle which ensure its proper
progression. Each checkpoint serves as a potential termination point along the cell cycle, during which
the conditions of the cell are assessed, with progression through the various phases of the cell cycle
occurring only when favorable conditions are met. There are many checkpoints in the cell cycle,[1] but
the three major ones are: the G1 checkpoint, also known as the Start or restriction checkpoint or Major
Checkpoint; the G2/M checkpoint; and the metaphase-to-anaphase transition, also known as the spindle
checkpoint.[2] Progression through these checkpoints is largely determined by the activation of cyclin-
dependent kinases by regulatory protein subunits called cyclins, different forms of which are produced
at each stage of the cell cycle to control the specific events that occur therein.[3][4]

Background[edit]

All living organisms are the products of repeated rounds of cell growth and division.[5] During this
process, known as the cell cycle, a cell duplicates its contents and then divides in two. The purpose of
the cell cycle is to accurately duplicate each organism's DNA and then divide the cell and its contents
evenly between the two resulting cells. In eukaryotes, the cell cycle consists of four main stages: G1,
during which a cell is metabolically active and continuously grows; S phase, during which DNA
replication takes place; G2, during which cell growth continues and the cell synthesizes various proteins
in preparation for division; and the M (mitosis) phase, during which the duplicated chromosomes
(known as the sister chromatids) separate into two daughter nuclei, and the cell divides into two
daughter cells, each with a full copy of DNA.[6] Compared to the eukaryotic cell cycle, the prokaryotic
cell cycle (known as binary fission) is relatively simple and quick: the chromosome replicates from the
origin of replication, a new membrane is assembled, and the cell wall forms a septum which divides the
cell into two.[7]

As the eukaryotic cell cycle is a complex process, eukaryotes have evolved a network of regulatory
proteins, known as the cell cycle control system, which monitors and dictates the progression of the cell
through the cell cycle.[5] This system acts like a timer, or a clock, which sets a fixed amount of time for
the cell to spend in each phase of the cell cycle, while at the same time it also responds to information
received from the processes it controls. The cell cycle checkpoints play an important role in the control
system by sensing defects that occur during essential processes such as DNA replication or chromosome
segregation, and inducing a cell cycle arrest in response until the defects are repaired.[8] The main
mechanism of action of the cell cycle checkpoints is through the regulation of the activities of a family of
protein kinases known as the cyclin-dependent kinases (CDKs), which bind to different classes of
regulator proteins known as cyclins, with specific cyclin-CDK complexes being formed and activated at
different phases of the cell cycle. Those complexes, in turn, activate different downstream targets to
promote or prevent cell cycle progression.[9]

G1 (restriction) checkpoint[edit]

Main article: Restriction point

The G1 checkpoint, also known as the restriction point in mammalian cells and the start point in yeast, is
the point at which the cell becomes committed to entering the cell cycle. As the cell progresses through
G1, depending on internal and external conditions, it can either delay G1, enter a quiescent state known
as G0, or proceed past the restriction point.[5] DNA damage is the main indication for a cell to "restrict"
and not enter the cell cycle. The decision to commit to a new round of cell division occurs when the cell
activates cyclin-CDK-dependent transcription which promotes entry into S phase. This check point
ensures the further process.[10]

During early G1, there are three transcriptional repressors, known as pocket proteins, that bind to E2F
transcription factors. The E2F gene family is a group of transcription factors that target many genes that
are important for control of the cell cycle, including cyclins, CDKs, checkpoint regulators, and DNA repair
proteins. Misregulation of the E2F family is often found in cancer cases, providing evidence that the E2F
family is essential for the tight regulation of DNA replication and division.[10] The three pocket proteins
are Retinoblastoma (Rb), p107, and p130, which bind to the E2F transcription factors to prevent
progression past the G1 checkpoint.

The E2F gene family contains some proteins with activator mechanisms and some proteins with
repressing mechanisms. P107 and p130 act as co-repressors for E2F 4 and E2F 5, which work to repress
transcription of G1-to-S promoting factors. The third pocket protein, Rb, binds to and represses E2F 1,
E2F 2, and E2F 3, which are the E2F proteins with activating abilities.[10]

Positive feedback plays an essential role in regulating the progression from G1 to S phase, particularly
involving the phosphorylation of Rb by a Cyclin/CDK protein complex. Rb without a phosphate, or
unphosphorylated Rb, regulates G0 cell cycle exit and differentiation. During the beginning of the G1
phase, growth factors and DNA damage signal for the rise of cyclin D levels, which then binds to Cdk4
and Cdk6 to form the CyclinD:Cdk4/6 complex.[11] This complex is known to inactivate Rb by
phosphorylation. However, the details of Rb phosphorylation are quite complex and specific compared
to previous knowledge about the G1checkpoint. CyclinD:Cdk4/6 places only one phosphate, or
monophosphorylates, Rb at one of its fourteen accessible and unique phosphorylation sites. Each of the
fourteen specific mono-phosphorylated isoforms has a differential binding preference to E2F family
members, which likely adds to the diversity of cellular processes within the mammalian body.[11]

E2F 4 and E2F 5 are dependent on p107 and p130 to maintain their nuclear localization. However, Cyclin
D:Cdk 4/6 also phosphorylates p107 and p130, a process which releases their bind from E2F 4 and 5
(which then escape to the cytoplasm), and allowing for E2F 1-3 to bind to the DNA and initiate
transcription of Cyclin E.[10] Rb proteins maintain their mono-phosphorylated state during early G1
phase, while Cyclin E is accumulating and binding to Cdk2.

CyclinE:Cdk2 plays an additional important phosphorylation role in the G1-to-S transition. Particularly,
CyclinE:Cdk2 promotes a positive feedback loop which creates and “all or nothing” switch. In many
genetic control networks, positive feedback ensures that cells do not slip back and forth between cell
cycle phases [12] Cyclin E:Cdk2 proceeds to phosphorylate Rb at all of its phosphorylation sites, also
termed “hyper-phosphorylate”, which ensures complete inactivation of Rb. The hyper phosphorylation
of Rb is considered the late G1 restriction point, after which the cell cannot go backwards in the cell
cycle. At this point, E2F 1-3 proteins bind to DNA and transcribe Cyclin A and Cdc 6.[11]

Cyclin-dependent kinase inhibitor 1B (CDKN1B), also known as p27, binds to and prevents the activation
of CyclinE:Cdk2 by inhibition. However, as Cyclin A accumulates and binds to Cdk2, they form and
complex and inhibit p27. The G1 phase cyclin-dependent kinase works together with S phase cyclin-
dependent kinase target p27 for degradation. In turn, this allows for full activation of Cyclin A:Cdk2, a
complex which phosphorylates E2F 1-3 initiating their disassociation from the DNA promoter sites. This
allows E2F 6-8 to bind to the DNA and inhibit transcription.[10] The negative feedback loop used to
successfully inhibit the inhibitor, p27, is another essential process used by cells to ensure mono-
directional movement and no backtrack through the cell cycle.

When DNA damage occurs, or when the cell detects any defects which necessitate it to delay or halt the
cell cycle in G1, arrest occurs through several mechanisms. The rapid response involves phosphorylation
events that initiate with either kinase ATM (Ataxia telangiectasia mutated) or ATR (Ataxia Telangiectasia
and Rad3 related), which act as sensors, depending on the type of damage. These kinases phosphorylate
and activate the effector kinases Chk2 and Chk1, respectively, which in turn phosphorylate the
phosphatase Cdc25A, thus marking it for ubiquitination and degradation. As Cdc25A activates the
previously mentioned cyclin E-CDK2 complex by removing inhibitory phosphates from CDK2, in the
absence of Cdc25A, cyclin E-CDK2 remains inactive, and the cell remains in G1.

To maintain the arrest, another response is initiated, by which Chk2 or Chk1 phosphorylate p53, a tumor
suppressor, and this stabilizes p53 by preventing it from binding Mdm2, a ubiquitin ligase which inhibits
p53 by targeting it for degradation. The stable p53 then acts a transcriptional activator of several target
genes, including p21, an inhibitor of the G1-to-S promoting complex cyclin E-CDK2. In addition, another
mechanism by which p21 is activated is through the accumulation of p16 in response to DNA damage.
p16 disrupts cyclin D-CDK4 complexes, thus causing the release of p21 from the complexes, which leads
to the dephosphorylation and activation of Rb, which allows Rb to bind and inhibit E2F 1-3, thus keeping
the cell from transitioning to S phase.[13] Recently, some aspects of this model have been disputed.[14]

G2 checkpoint[edit]

See also: G2-M DNA damage checkpoint

Mitotic Cyclin Concentration shows hysteresis and bistability relative to Cdk1 Activation

Following DNA replication in S phase, the cell undergoes a growth phase known as G2. During this time,
necessary mitotic proteins are produced and the cell is once more subjected to regulatory mechanisms
to ensure proper status for entry into the proliferative Mitotic (M) phase. Multiple mechanistic
checkpoints are involved in this transition from G2 to M, with a common uniting factor of cyclin-Cdk
activity.

Although variations in requisite cyclin-Cdk complexes exist across organisms, the necessity of the kinase
activity is conserved and typically focuses on a single pairing. In fission yeast three different forms of
mitotic cyclin exist, and six in budding yeast, yet the primary cyclin utilized is cyclin B.[15] Cyclin B will
serve as reference for discussion of the G2/M checkpoint transition.

Similar to S Phase, G2 experiences a DNA damage checkpoint. The cell is once more examined for sites
of DNA damage or incomplete replication, and the kinases ATR and ATM recruited to damage sites.
Activation of Chk1 and Chk2 also transpire, as well as p53 activation, to induce cell cycle arrest and halt
progression into mitosis. An additional component of S phase, the Pre-Replicative Complex, must be
inactivated via cyclin B-Cdk1 phosphorylation.[16]

As these previous checkpoints are assessed, G2 protein accumulation serves to activate cyclinB-Cdk1
activity via multiple mechanisms. CyclinA-Cdk2 activates Cdc25, an activator of cyclinB-Cdk1, which then
deactivates the cyclinB-Cdk1 inhibitor, Wee1. This results in a positive feedback loop, significantly
increasing cyclinB expression and Cdk1 activation. As the cell progresses through G2 and reaches the
G2/M transition, the kinase Plk1 phosphorylates Wee1, which targets Wee1 for degradation via the SCF
ubiquitin ligase complex.[17] An additional function of Plk1 is to activate Cdc25 through
phosphorylation. The compound effect of Wee1 degradation and Cdc25 activation is the net removal of
inhibitory phosphorylation from cdc2, which activates cdc2. Plk1 is activated at the G2/M transition by
the Aurora A and Bora, which accumulate during G2 and form an activation complex. The Plk1-Cdc2-
cdc25 complex then initiates a positive feedback loop which serves to further activate Cdc2, and in
conjunction with an increase in cyclin B levels during G2, the resulting cdc2-cyclin B complexes then
activate downstream targets which promote entry into mitosis.[18] The resultant Cdk1 activity also
activates expression of Mem1-Fkh, a G2/M transition gene.[19] The rapid surge in cyclinB-Cdk1 activity
is necessary, as M phase initiation is an all-or-nothing event engaging in hysteresis. Hysteresis of Cdk1
activity via cyclin B drives M phase entry by establishing a minimum threshold of cyclinB concentration.
This exists at a level higher than the minimum needed for the continuation of M phase after entry,
acting to safeguard the all-or-nothing event. This entry concentration is further increased in the case of
incomplete DNA replication, adding another regulatory mechanism at the G2/M transition point.[20]
The presence of hysteresis allows for M phase entry to be highly regulated as a function of cyclinB-Cdk1
activity.

The mechanisms by which mitotic entry is prevented in response to DNA damage are similar to those in
the G1/S checkpoint. DNA damage triggers the activation of the aforementioned ATM/ATR pathway, in
which ATM/ATR phosphorylate and activate the Chk1/Chk2 checkpoint kinases. Chk1/2 phosphorylate
cdc25 which, in addition to being inhibited, is also sequestered in the cytoplasm by the 14-3-3 proteins.
14-3-3 are upregulated by p53, which, as previously mentioned, is activated by Chk1 and ATM/ATR. p53
also transactivates p21, and both p21 and the 14-3-3 in turn inhibit cyclin B-cdc2 complexes through the
phosphorylation and cytoplasmic sequestering of cdc2. In addition, the inactivation of cdc25 results in its
inability to dephosphorylate and activate cdc2.[21][22] Finally, another mechanism of damage response
is through the negative regulation of Plk1 by ATM/ATR, which in turn results in the stabilization of Wee1
and Myt1, which can then phosphorylate and inhibit cdc2, thus keeping the cell arrested in G2 until the
damage is fixed.[23]

Metaphase checkpoint[edit]

Main article: Spindle checkpoint

The mitotic spindle checkpoint occurs at the point in metaphase where all the chromosomes
should/have aligned at the mitotic plate and be under bipolar tension. The tension created by this
bipolar attachment is what is sensed, which initiates the anaphase entry. To do this, the sensing
mechanism ensures that the anaphase-promoting complex (APC/C) is no longer inhibited, which is now
free to degrade cyclin B, which harbors a D-box (destruction box), and to break down securin.[24] The
latter is a protein whose function is to inhibit separase, which in turn cuts the cohesins, the protein
composite responsible for cohesion of sister chromatids.[25] Once this inhibitory protein is degraded via
ubiquitination and subsequent proteolysis, separase then causes sister chromatid separation.[26] After
the cell has split into its two daughter cells, the cell enters G1.
 M phase

During the mitotic (M) phase, the cell divides its copied DNA and cytoplasm to make two new
cells. M phase involves two distinct division-related processes: mitosis and cytokinesis.

In mitosis, the nuclear DNA of the cell condenses into visible chromosomes and is pulled apart
by the mitotic spindle, a specialized structure made out of microtubules. Mitosis takes place in four
stages: prophase (sometimes divided into early prophase and prometaphase), metaphase, anaphase,
and telophase. You can learn more about these stages in the video on mitosis.

In cytokinesis, the cytoplasm of the cell is split in two, making two new cells. Cytokinesis usually
begins just as mitosis is ending, with a little overlap. Importantly, cytokinesis takes place differently in
animal and plant cells

n animals, cell division occurs when a band of cytoskeletal fibers called the contractile ring
contracts inward and pinches the cell in two, a process called contractile cytokinesis. The indentation
produced as the ring contracts inward is called the cleavage furrow. Animal cells can be pinched in two
because they’re relatively soft and squishy.
 Plant cells are much stiffer than animal cells; they’re surrounded by a rigid cell wall and have
high internal pressure. Because of this, plant cells divide in two by building a new structure down the
middle of the cell. This structure, known as the cell plate, is made up of plasma membrane and cell wall
components delivered in vesicles, and it partitions the cell in two.

Cell cycle exit and G_0

start subscript, 0, end subscript

What happens to the two daughter cells produced in one round of the cell cycle? This depends
on what type of cells they are. Some types of cells divide rapidly, and in these cases, the daughter cells
may immediately undergo another round of cell division. For instance, many cell types in an early
embryo divide rapidly, and so do cells in a tumor.

Other types of cells divide slowly or not at all. These cells may exit the G_1

start subscript, 1, end subscript phase and enter a resting state called G_0

start subscript, 0, end subscript phase. In G_0

start subscript, 0, end subscript, a cell is not actively preparing to divide, it’s just doing its job.
For instance, it might conduct signals as a neuron (like the one in the drawing below) or store
carbohydrates as a liver cell. G_0

start subscript, 0, end subscript is a permanent state for some cells, while others may re-start
division if they get the right signals.
 e events of Meiosis II are analogous to those of a mitotic division, although the number of
chromosomes involved has been halved.

Meiosis generates genetic diversity through:

the exchange of genetic material between homologous chromosomes during Meiosis I

the random alignment of maternal and paternal chromosomes in Meiosis I

the random alignment of the sister chromatids at Meiosis II

Regulator Molecules of the Cell Cycle

In addition to the internally controlled checkpoints, there are two groups of intracellular
molecules that regulate the cell cycle. These regulatory molecules either promote progress of the cell to
the next phase (positive regulation) or halt the cycle (negative regulation). Regulator molecules may act
individually or they can influence the activity or production of other regulatory proteins. Therefore, the
failure of a single regulator may have almost no effect on the cell cycle, especially if more than one
mechanism controls the same event. Conversely, the effect of a deficient or non-functioning regulator
can be wide-ranging and possibly fatal to the cell if multiple processes are affected.

Positive Regulation of the Cell Cycle

Two groups of proteins, called cyclins and cyclin-dependent kinases (Cdks), are responsible for
the progress of the cell through the various checkpoints. The levels of the four cyclin proteins fluctuate
throughout the cell cycle in a predictable pattern. Increases in the concentration of cyclin proteins are
triggered by both external and internal signals. After the cell moves to the next stage of the cell cycle,
the cyclins that were active in the previous stage are degraded.

Cyclins regulate the cell cycle only when they are tightly bound to Cdks. To be fully active, the
Cdk/cyclin complex must also be phosphorylated in specific locations. Like all kinases, Cdks are enzymes
(kinases) that phosphorylate other proteins. Phosphorylation activates the protein by changing its
shape. The proteins phosphorylated by Cdks are involved in advancing the cell to the next phase.. The
levels of Cdk proteins are relatively stable throughout the cell cycle; however, the concentrations of
cyclin fluctuate and determine when Cdk/cyclin complexes form. The different cyclins and Cdks bind at
specific points in the cell cycle and thus regulate different checkpoints.
 Although the cyclins are the main regulatory molecules that determine the forward momentum
of the cell cycle, there are several other mechanisms that fine tune the progress of the cycle with
negative, rather than positive, effects. These mechanisms essentially block the progression of the cell
cycle until problematic conditions are resolved. Molecules that prevent the full activation of Cdks are
called Cdk inhibitors. Many of these inhibitor molecules directly or indirectly monitor a particular cell
cycle event. The block placed on Cdks by inhibitor molecules will not be removed until the specific event
being monitored is completed.

Negative Regulation of the Cell Cycle

The second group of cell cycle regulatory molecules are negative regulators. Negative regulators
halt the cell cycle. Remember that in positive regulation, active molecules cause the cycle to progress.

The best understood negative regulatory molecules are retinoblastoma protein (Rb), p53, and
p21. Retinoblastoma proteins are a group of tumor-suppressor proteins common in many cells. Much of
what is known about cell cycle regulation comes from research conducted with cells that have lost
regulatory control. All three of these regulatory proteins were discovered to be damaged or non-
functional in cells that had begun to replicate uncontrollably (became cancerous). In each case, the main
cause of the unchecked progress through the cell cycle was a faulty copy of the regulatory protein.

Rb, p53, and p21 act primarily at the G1 checkpoint. p53 is a multi-functional protein that has a
major impact on the cell’s commitment to division; it acts when there is damaged DNA in cells that are
undergoing the preparatory processes during G1. If damaged DNA is detected, p53 halts the cell cycle
and recruits enzymes to repair the DNA. If the DNA cannot be repaired, p53 can trigger apoptosis (cell
suicide) to prevent the duplication of damaged chromosomes. As p53 levels rise, the production of p21
is triggered. p21 enforces the halt in the cycle dictated by p53 by binding to and inhibiting the activity of
the Cdk/cyclin complexes. As a cell is exposed to more stress, higher levels of p53 and p21 accumulate,
making it less likely that the cell will move into the S phase.

Rb exerts its regulatory influence on other positive regulator proteins. Rb monitors cell size. In
the active, dephosphorylated state, Rb binds to proteins called transcription factors, most commonly to
E2F. Transcription factors “turn on” specific genes, allowing the production of proteins encoded by that
gene. When Rb is bound to E2F, production of proteins necessary for the G1/S transition is blocked. As
the cell increases in size, Rb is slowly phosphorylated until it becomes inactivated. Rb releases E2F,
which can now turn on the gene that produces the transition protein and this particular block is
removed. For the cell to move past each of the checkpoints, all positive regulators must be “turned on”
and all negative regulators must be “turned off.”