The Cell Cycle Control SystemOverview

The cell cycle is an organized set of events that leads the cell to divide into two daughter
cells, each containing chromosomes identical to the parent cell. It is the cell cycle that leads
to the formation of an entire organism from a single-cell zygote. Besides, cell division also
functions in the renewal or repair of tissues in adult multicellular eukaryotes. For example,
in the bone marrow, the stem cells divide to form new blood cells. Although essential for
several functions, cell division in the absence of a control mechanism leads to cancer and
many genetic diseases.

To ensure DNA replication occurs correctly and each daughter cell inherits the right number
of chromosomes, the cell has surveillance mechanisms that make up the cell cycle control
system. There are at least two known cell cycle control methods. One of these processes
includes a cascade of protein phosphorylations that transitions a cell from one phase to the
next. Also, there is a series of checkpoints that monitor the completion of essential events
and, if necessary, delay progression to the next phase. At every checkpoint, the regulator
proteins prevent the cell initiation from entering the next phase until the previous phase's
errors are rectified.

The first form of regulation includes a highly regulated family of kinases. Kinase activation
requires interaction with a second subunit expressed at fixed points of the cell cycle. This
secondary component is termed as- a phase-specific "cyclin"  that combines with its partner
"cyclin-dependent kinase" (CDK), forming an active complex, each of which exhibits distinct
substrate specificity. Regulatory phosphorylation and dephosphorylation fine-tune the
function of cyclin-CDK complexes, ensuring a well-defined progression.

The second form of cell cycle regulation - checkpoint control, is more of a surveillance
mechanism. Cell cycle checkpoints identify defects in crucial events such as DNA
replication and chromosome segregation. For instance, DNA damage triggers a signaling
cascade that activates several cell cycle inhibitors. These inhibitors bind critical cell cycle
proteins to arrest the cycle until the risk of mutation has been eliminated.

Procedure

The cell cycle involves the duplication of the intracellular content, followed by the division
into two daughter cells.

Replication of cellular content, especially the DNA, is highly critical, and any mistake during
the process can lead to conditions such as cancer. 

So how do cells prevent any errors during division?

The cell cycle control system features regulator proteins that halt the cycle at various
checkpoints.

At every checkpoint, the regulator proteins prevent the initiation of each step until the
earlier stages are completed and any errors have been corrected.

Generally, the control system has three crucial checkpoints found in the G1, G2, and M
phases.
At the G1 checkpoint, the regulator protein checks if the cell has reached the critical size
and the DNA is free from errors. It also checks whether enough nutrients and growth
factors are present to begin DNA synthesis.

At this point, if the cell does not receive the necessary signal, it switches to a resting state
called the G0 phase until all the conditions are met.

The cells that pass the G1 checkpoint transit through the synthesis, or S-phase, when the
DNA gets replicated.

After this, the cell encounters a second checkpoint at the G2 phase, where the regulator
proteins check any errors in the DNA and whether the cell has an appropriate size to enter
Mitosis or M-phase.

During Mitosis, the control system verifies if the chromosomes are attached to the spindle
and are accurately aligned so that the cycle proceeds for cell division. If at any point the
regulator protein detects irreparable damage, cell death occurs. One type of critical
regulator proteins are the cyclin-dependent protein kinases, or CDK. CDKs form complexes
with cyclins, and their activity affects proteins directly involved in cell growth and DNA
synthesis.

Positive Regulator Molecules

Overview

Mitotic cell division results in daughter cells that exactly resemble the parent cell. However,
errors in the DNA replication or distribution of genetic material may lead to genetic
mutations that may be passed down to every new cell formed from the resulting abnormal
cell. Propagation of such mutant cells is restricted through checkpoint mechanisms present
at different stages of the cell cycle. These checkpoints involve regulator molecules that
either promote or demote cell cycle events.

Proteins, such as cyclin and cyclin-dependent kinases, are positive regulator molecules
responsible for the cell cycle’s progression through various checkpoints. The cyclins were
initially named such because their synthesis and degradation assume a cyclical pattern.
There are at least four functional cyclins whose concentration fluctuates predictably across
the cell cycle. When a cell gets promoted to the next stage, the cyclins of the previous
phase get degraded. It is the changes in the cyclin concentration that trigger various cell
cycle events.

Cyclins form an active complex with protein kinases that can phosphorylate specific target
proteins during the cell cycle. Because these kinases need cyclin for activation, they are
called cyclin-dependent kinases or CDKs. In the absence of cyclin, the CDKs are inactive,
and in the absence of a fully activated cyclin/CDK complex, the cell fails to pass through
the checkpoints.

The positive regulator molecules are expressed by genes that belong to a group called
proto-oncogenes. When mutated, these become oncogenes that cause the cell to become
cancerous. For instance, a mutation that causes the CDKs to become active even in the
absence of cyclin can cause the mutant cell to pass uninterrupted through the checkpoints
leading to uncontrolled growth and proliferation.

Procedure

The positive regulator molecules promote the transition through various stages of the cell
cycle via the interaction of two protein groups: cyclins and cyclin-dependent kinases or
Cdks.

Mammalian cells contain around nine Cdks, with four of them, Cdk1, Cdk6, Cdk4, and
Cdk2, involved in the cell cycle.

The activity and specificity of any given Cdk depends on the binding of cyclins. Cyclins are
grouped as G1, G1/S, S, or M phase cyclins, and their expression is specific to the stage
they trigger

For instance, in the G1 phase, cyclin D binds to Cdk4 and Cdk6, promoting the cell to the
late G1 phase.

Next, cyclin E accumulates and forms a complex with Cdk2. The cyclin E-Cdk2 complex,
along with the cyclin D-Cdk4/6, triggers the G1 to S transition, which irreversibly commits
the cells into the cycle.

At the start of S phase, cyclin-E remains elevated and bound to Cdk2. In addition, cyclin-A
level rises and combines with Cdk2. Both the complexes are directly responsible for DNA
replication.

Although cyclin E levels fall, cyclin-A levels are high throughout the S and G2-phases. At
the G2-M transition, cyclin A levels drop and cyclin B levels rise.

Cyclin B combines with Cdk1, triggering the start of mitosis. The levels of mitotic cyclins fall
mid-mitosis, thus inactivating the Cdk.

The inactive Cdk has a protein loop that blocks substrate protein access to the active site. It
is only when a specific cyclin binds to the Cdk, that the loop moves away from the active
site, thus partially activating Cdk.

The complete activation of the Cdk-cyclin complex depends on another enzyme called Cdk
activating kinase or CAK.

The CAK phosphorylates an amino acid near the active site causing a conformational
change in Cdk, enabling the CDK-cyclin complex to phosphorylate its target proteins and
induce specific cell cycle stage events.

Inhibition of CDK Activity

Overview

The orderly progression of the cell cycle depends on the activation of CDK protein by
binding to its cyclin partner. However, the cell cycle must be restricted when the cell
undergoes abnormal changes. Most cancers correlate to the deregulated cell cycle, and
since CDKs are a central component of the cell cycle, the CDK inhibitors are extensively
studied to develop anticancer agents. For instance, cyclin D associates with several CDKs,
such as CDK 4/6, to form an active complex. The cyclin D-CDK4/6 complex then
phosphorylates and inactivates the tumor suppressor retinoblastoma protein (Rb) to
promote the G1-to-S phase transition of the cell cycle. In normal cells, Rb protein is
reactivated through the regulation of CDK activity, thus, preventing abnormal cell cycle
transitions.

There are at least three known mechanisms by which CDK activity is regulated- cyclin
degradation, inhibitory phosphorylation, or binding of inhibitory proteins. Mutations that
prevent these mechanisms lead to CDK-mediated tumorigenesis.

Because CDK 4/6 plays a substantial role in tumor formation, several CDK inhibitors have
been developed for clinical use. The most recent ones are selective for CDK4 and CDK6.
There are at least three clinically approved CDK 4/6 inhibitors: abemaciclib, ribociclib, and
palbociclib. These inhibitors bind to the ATP pocket of CDK 4 and 6, inactivating the Cyclin
D-CDK4/6 complexes, leading to Rb protein activation and subsequent cell cycle arrest. In
some cases, the inhibitor-mediated  cell cycle arrest causes an increase in apoptosis in
tumor cells.

Inhibition of the cell cycle and subsequent programmed cell death are the most common
mechanisms of CDK4/6 inhibitors. However, a recent study in mouse models of breast
cancer showed that CDK4/6 inhibition could also lead to severe immunogenic effects.
During the study, the CDK inhibitor seemed to enhance tumor cells' antigen-presenting
ability, thereby allowing cytotoxic T cells to recognize and destroy the tumor cells.

Procedure

During cell cycle transitions, Cdk activity is regulated by multiple proteins, ensuring
controlled cell growth, complete DNA replication, and mitotic distribution of the
chromosomes to daughter cells.

In the absence of regulatory proteins, an abnormal cell goes unchecked, leading to

conditions such as cancer.

In a normal cell, Cdk activity is regulated through multiple mechanisms including cyclin
degradation, inhibitory phosphorylation, and inhibitory conformational changes induced by
Cdk inhibitor binding.

Cyclin levels are known to fluctuate during the cell cycle. Cdks are only active when bound
to cyclin, therefore, cyclin degradation leaves Cdks inactive and unable to promote the
transition to the next cell cycle stage.

In another mechanism, a kinase named Wee1, phosphorylates the active site in Cdk.  This
phosphorylation inhibits the activity of the cyclin-Cdk complex.

Additionally, Cdk inhibitors or CKIs, such as p16, p21, and p27, regulate Cdk activity
through inhibitory conformational changes.
For instance, if DNA damage occurs during G1, p16 interacts with the cyclin-Cdk complex.
This interaction causes a large structural rearrangement, detaching the bound cyclin,
causing Cdk inactivation. 

DNA damage in the G1 phase can also trigger the tumor suppressor protein, p53, to
activate p21.

G1/S-Cdk and S-Cdk complexes are bound and inhibited by p21, arresting the cells in the
G1 phase, and allowing enough time for DNA repair.

For the G1/S phase transition to take place, the cell is required to have all the resources
needed for DNA replication. If the cell lacks the necessary resources, p27 binds to G1/S-
Cdk and S-Cdk complexes, inhibiting the enzyme activity.

Once favorable conditions for cell cycle progression are met, p27 is degraded, restoring
Cdk activity and promoting cell transition.

S-Cdk Initiates DNA Replication

Overview

The cell cycle is a series of events leading to DNA duplication followed by the division of
cell content to form two daughter cells. The cell cycle progresses in four stages – the cell
increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares
to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).

Two states at the origin of replication

In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called
the origins of replication. During the progression of the cell cycle, the origins of replication
exist in two states. The first state exists in the G1-phase when a multiprotein complex
called the pre-replicative complex (pre-RC) assembles on the origin. The second state
exists from the initiation of the S-phase to the end of the M-phase when a complex with
fewer components called the post-replicative complex (post-RC) remains on the origin
DNA.

CDK activity controls each round of DNA replication

At the end of the M-phase, cyclin-dependent kinase (CDK) activity is low within the cells,
which permits the pre-RC assembly, resulting in a replication-competent state. During the
G1–S phase transition, CDK activity increases, triggering DNA replication initiation. The
increased CDK activity also causes the disassembly of the pre-RC complex, converting the
origin to a post-RC state. Throughout the S-phase and the remainder of the cell cycle,
persistent high CDK activity prevents the re-assembly of pre-RC until the end of mitosis
when the CDK activity again reduces. This control mechanism inhibits re-replication.

Procedure
In eukaryotes, DNA replication takes place during the S-phase of the cell cycle. The cell
cycle control system, consisting of a network of regulatory proteins, governs the
progression of the cell cycle.

The S-phase cyclin-dependent kinase, or SCDKs, are enzyme complexes involved in cell
cycle control during S-phase. This control system ensures that every nucleotide in the
genome is copied only once during replication.

DNA replication begins at the origins of replication, present at multiple locations in the
chromosomes. Before the initiation of replication, multiprotein complexes called origin
recognition complexes bind to the DNA and serve as docking sites for other proteins.

During early G1-phase of the cell cycle, the regulatory proteins Cdc6 and Cdt1 bind to the
ORCs, and aid in the assembly of a set of proteins called the MCM proteins into inactive
ring complexes on the adjacent DNA, thereby forming large multiprotein complexes called
the pre-replicative complexes or pre-RCs. MCMs function as DNA helicases for replication.

At the onset of S-phase, S-CDKs are activated and trigger origin firing, or initiation of DNA
replication, by phosphorylating specific initiator proteins. Phosphorylated initiator proteins
promote the recruitment of helicase activator complexes. These complexes activate the
DNA helicases and recruit the DNA polymerase, leading to replication.

S-CDKs not only initiate replication but also prevent re-replication from occurring at the
same origin.

S-CDKs phosphorylate the Cdc6 and Cdt1 proteins, promoting their release from the
ORCs, leading to degradation, and the disassembly of the pre-RCs.

After DNA replication, when the helicases disengage from the DNA strand, the S-CDKs
phosphorylate the helicases, triggering their export from the nucleus.

This cell cycle control mechanism involving S-CDKs prevents re-initiation of replication,
thereby ensuring that the DNA replication occurs only once per cell cycle.

M-Cdk Drives Transition Into Mitosis

Overview

Checkpoints throughout the cell cycle serve as safeguards and gatekeepers, allowing the
cell cycle to progress in favorable conditions and slow or halt it in problematic ones. This
regulation is known as the cell cycle control system.

Cyclin-dependent kinases, or CDKs, work in concert with cyclins to control cell cycle
transitions. M-CDK, a complex of CDK1 bound to M cyclin, is a well-known example of this
coordinated control that drives the transition from G2 to M phase.

M cyclin promotes M phase events—such as mitotic spindle formation, sister chromatid

attachment to opposite spindle poles, chromosome condensation, nuclear envelope
breakdown, and actin cytoskeleton and Golgi apparatus rearrangement. By promoting
these processes, M cyclin drives the transition into the M phase.
Like other cyclins, M cyclin levels fluctuate during the cell cycle. CDK levels, however,
remain relatively stable. In most cells (embryonic cells are an exception), M cyclin gene
transcription increases, and M cyclin accumulates as the cell approaches the G2/M
transition. The accumulated M cyclin binds to CDK, forming M-CDK complexes. The M-
CDKs are primed to trigger M phase events upon activation, largely by Cdc25.

Active M-CDK enables the transition into mitosis by phosphorylating proteins that enable
several early mitotic processes. However, M-CDK is not the only protein kinase that
regulates this transition. Polo-like kinases and Aurora kinases, for example, also contribute
to early mitotic processes.

Plk1 is a polo-like kinase necessary for the normal bipolar formation of the mitotic spindle.
Plk1 phosphorylates proteins that help separate the spindle poles. Aurora-A also regulates
proteins involved in forming and stabilizing the mitotic spindle, while Aurora-B allows sister
chromatids to attach to the spindle. Together, M-CDK and other protein kinases help
regulate the transition between the G2 and M phases of the cell cycle.

Procedure

A cell’s transition into mitosis is characterized by the activation of M-CDK complexes,

consisting of the protein kinase CDK1—or cyclin-dependent kinase 1—bound to M cyclin.

M-CDK complexes form when M cyclin accumulates. In most cells, M cyclin levels peak
during G2—the gap phase following the chromosomal duplication of S phase—and early
mitosis.

The M-CDK complex is phosphorylated at an active site by CDK-activating kinase, or CAK.

However, the complex remains inactive, because it is also phosphorylated at two inhibitory
sites by the protein kinase Wee1.

M-CDK is activated largely by the protein phosphatase Cdc25. Cdc25 removes the
phosphates that inhibit M-CDK and suppresses the inhibitory activity of Wee1.

M-CDK drives the transition into mitosis by activating factors necessary for early mitotic
processes.

In prophase, M-CDK activity spurs the shortening and compaction of chromosomes, known
as chromosome condensation. During prophase, M-CDK also initiates the formation of the
mitotic spindle—which separates chromosomes into two daughter cells.

During prometaphase in animal cells, M-CDK helps degrade the nuclear envelope, allowing
the nucleus to break apart.

In metaphase, M-CDK mediates the attachment of sister chromatids to opposite poles of

the spindle.

M-CDK promotes the multiphase reorganization of the Golgi apparatus, which is important
for correct spindle formation and segregation of the organelle. In addition, throughout
mitosis,  M-CDK is involved in the reorganization of the actin cytoskeleton, which helps
determine spindle orientation and the axis of cell division, among other functions.
Mitogens and the Cell CycleOverview

Mitogens and their receptors play a crucial role in controlling the progression of the cell
cycle. However, the loss of mitogenic control over cell division leads to tumor formation.
Therefore, mitogens and mitogen receptors play an important role in cancer research. For
instance, the epidermal growth factor (EGF) - a type of mitogen and its transmembrane
receptor (EGFR), decides the fate of the cell's proliferation. When EGF binds to EGFR, a
member of the ErbB family of tyrosine kinase receptors present on the cell membrane, it
transmits a growth-inducing signal to the corresponding cells. However, overactivation of
EGFR can lead to tumor growth, invasion, and metastasis. It needs to be inactivated in the
cancer cells to induce cell cycle arrest, dedifferentiation, or programmed cell death. Hence,
the development of novel and targeted cancer therapies requires a deeper understanding
of the mechanism and coordination between the mitogen and the cell cycle.

Role of Epidermal Growth Factor (EGF) as a mitogen in cancer therapeutics

In non-malignant tissues, the number of EGFR on the cell surface is tightly regulated to
ensure that the cell division rate accurately matches the tissue's requirement. However, in
cancerous cells, EGFR is overexpressed and is often perpetually stimulated by EGF or
EGF-like proteins secreted by the cancer cell itself. A similar effect may occur when a
mutation in EGFR drives the receptor into a state of continual activation. Overexpression of
EGFR and closely associated ErbB2 receptors are associated with more aggressive clinical
behavior, such as in Grade 3 cancers where tumor cells can likely spread to other parts of
the body. Therefore, therapies directed at inhibiting the function of overactive receptors in
the cancer cells can be used as anti-cancer therapies.

Monoclonal antibodies (MAbs) that block activation of EGFR and ErbB2 have been
developed. These MAbs have shown promising preclinical studies. For example,
trastuzumab, an anti-ErbB2 MAb, was recently approved to treat patients with metastatic
ErbB2-overexpressing breast cancer. Another MAb, IMC-C225, an anti-EGFR, has shown
impressive activity to revert tumor cells' resistance to chemotherapy.

Procedure

Unicellular organisms, such as bacteria, divide when there are enough nutrients in the
environment.

However, in multicellular organisms, most cells remain in G0, or the non-dividing phase,
until cell division is triggered by extracellular signal molecules, called 'mitogens’.

Mitogens are usually small proteins or peptides secreted in response to different stimuli,
such as tissue injury, infection, or a routine regeneration.

For instance, in the case of tissue injury, specialized cells secrete a mitogen called Platelet-
derived growth factor or PDGF.

Such mitogens can bind to the extracellular domain of tyrosine kinase receptors such as
PDGF-Receptor, inducing receptor dimerization and autophosphorylation of its intracellular
domains. This allows phosphorylation and activation of the downstream intracellular
molecules that are involved in multiple signaling pathways.
The mitogen-activated protein kinase or MAP kinase cascades are a type of signaling
pathway induced by mitogen-receptor binding. This pathway starts with the activation of the
small membrane bound GTPase named Ras by the receptor.

Next, active Ras triggers the MAP kinase cascade which includes a series of kinase
proteins. In a chain of phosphorylation reactions, first MAP3 kinase activates MAP2 kinase,
and then finally MAP kinase.

Active MAPK then translocates into the nucleus where it activates regulatory transcription
factors, including Myc.

Myc increases the expression of G1 cyclins that partner with cyclin-dependent kinases or
Cdks.

The G1 cyclin-Cdk complex phosphorylate Rb - a tumor suppressor protein - freeing the

bound gene regulatory factor-E2F. Active E2F then binds to a specific DNA sequence and
triggers transcription of cell cycle genes that encode several proteins necessary for cell
division.

As the mitogenic stimuli recede, such as during the healing of injured tissues, Rb protein
interacts with E2F inhibiting its activity, thus, preventing excessive cell growth.

Replicative Cell Senescence

Overview

Replicative cell senescence is a property of cells that allows them to divide a finite number
of times throughout the organism's lifespan while preventing excessive proliferation.
Replicative senescence is associated with the gradual loss of the telomere — short,
repetitive DNA sequences found at the end of the chromosomes. Telomeres are bound by
a group of proteins to form a protective cap on the ends of chromosomes. Embryonic stem
cells express telomerase — an enzyme that adds the telomeric repeat sequence and
enables repetitive cell division; however, in adults, telomerase is active only in the cells that
need to divide regularly.

Because telomerase is inactive in most human somatic cells, the length of the telomere
decreases with every cell division. After a critical length, telomere shortening leads to
permanent cell cycle arrest. This mechanism is assumed to protect against cancer
development by limiting the abnormal proliferation of tumors; however, a rare mutation can
activate telomerase, which reconstructs the telomere region, allowing the cells to
proliferate. Thus, telomerase is a perfect target for specific anticancer therapy, as most
cancer cells express telomerase while normal cells do not.

The relation between telomere length and tumor formation has been experimentally verified
using oncogenic mice. Oncogenic mice are mouse models that have cancer-causing
genes. When such oncogenic mice are crossed with telomerase-deficient mice that lack
telomerase activity, the resultant progeny mice express shorter telomeres than the
oncogenic parent. These progeny mice, when interbred, generate successive generations
that have progressively shorter telomeres. The frequency of tumor formation is studied by
treating the progeny mice with carcinogens at every generation. As per the results, the late
generation mice with shorter telomeres exhibit a reduced frequency of tumor formation
compared to the early generation mice that maintain longer telomeres. This proves that
limiting the replicative capacity of cells suppresses tumor formation.

Procedure

Most animal cells divide a finite number of times before they stop and undergo permanent
cell cycle arrest.

In a mitogenic medium, for example, human fibroblast cells divide about 25-50 times. As a
cell approaches this finite number of divisions, the rate of cell division slows down and
finally halts with cells entering a permanent non-dividing state. This phenomenon is called
replicative cell senescence.

Replicative cell senescence is a result of changes in the structure of telomeres. Telomeres

are located at the ends of the chromosomes and consist of long repetitive DNA sequences
and protein complexes.

In the absence of telomeres, chromosome ends could be recognized as double-strand

breaks. These ends could fuse to one another forming abnormal structures like a ring
chromosome. The telomeres act as caps, protecting the ends of the chromosomes from
degradation by nucleases and preventing the aberrant fusion of chromosome ends to one
another.

Shelterin is a telomere-associated protein complex that protects the chromosome ends.

Shelterin helps DNA ends form a lariat-like structure called a telomerase-loop, or T-loop.
This T-loop masks the DNA ends, preventing degradation.

During cell division, telomeres are shortened by 25-200 bases due to the inability of the
polymerase to completely replicate DNA ends. As the length of telomeres becomes shorter,
the shelterin components are displaced from the telomere region. Shrinking of the telomere
eventually destabilizes the t-loop conformation.

The change in the T-loop structure leaves the chromosome ends exposed, which are
sensed as DNA damage by the DNA damage response pathway.

The persistent DNA damage response that ensues induces replicative cell senescence
which helps limit genomic instability and malignant transformation.

Abnormal Proliferation

Overview

Under normal conditions, most adult cells remain in a non-proliferative state unless
stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a
condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such
situations, cell division persists in the same excessive manner even after cessation of the
stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate
to pass the damage to the daughter cells and all the future daughter cells. Such abnormal
cell proliferation defines the characteristic of what we call cancer. The best example to
explain this condition is the mutation in the RAS and MYC proteins.

Normal RAS proteins are GTPases that function in signal transduction from cell surface
receptors to the cell’s interior. RAS proteins can be denoted as the binary switches that
cycle between ON and OFF states during cellular growth. Usually, these switches are
tightly regulated, but in RAS-related diseases such as cancer, the RAS gene mutation or
their regulator renders RAS proteins persistently active. For instance, a single amino acid
missense mutation in the KRAS protein (a type of RAS protein) impairs its normal
functioning. The mutant KRAS cannot be inactivated, leading to continuous growth
stimulation. KRAS mutation occurs in 15%- 20% of human cancers, and is most commonly
seen in colon cancer, lung cancer, pancreatic cancer, and leukemia.

Abnormal cell proliferation can also occur when mutation results in overexpression of MYC
protein. The MYC oncogene belongs to a family of "super-transcription factors" and is
deregulated in more than 50% of human cancers. The MYC affects a spectrum of cellular
functions, including protein translation, cell cycle progression, ribosome biosynthesis, cell
proliferation, differentiation, survival, and immune surveillance.

Procedure

 Cells have special genes called tumor suppressor genes that can neutralize the effect of
harmful genetic alterations and prevent uncontrolled cell proliferation.

For example, p53 is a tumor suppressor gene that maintains a basal level of expression
under normal cell conditions.

Mdm2 protein acts as a negative regulator of p53 activity. It binds to the functional domain
of p53 protein, reducing its transcriptional activity.

Mdm2 also exhibits p53-specific ubiquitin ligase activity and catalyzes the attachment of
ubiquitin to p53. The polyubiquitinated p53 proteins are then recognized and degraded by
the proteasome, maintaining it at extremely low levels inside the cell.

However, when the cells encounter abnormal conditions such as cellular stress or
excessive mitogenic stimulation, the p53 gene is induced to enforce its tumor suppression
activity.

For instance, overexpression of transcription factor Myc in the nucleolus triggers the
accumulation of a tumor suppressor protein called- Arf.

Arf directly binds to Mdm2, inhibiting its ubiquitin ligase activity and sequestering it in the
nucleolus, thus releasing active p53 into the nucleoplasm.

Active p53 then binds to specific DNA sequences and induces the expression of its target
genes that can trigger accelerated DNA repair, arrest cell cycle, or induce cell apoptosis,
thus, preventing further propagation of damaged cells.

 However, when genetic or epigenetic changes alter p53 activity, it results in the collapse of
the cell’s protective mechanisms. Such cells start proliferating aggressively and form
tumors.
Besides, overexpression of Mdm2 is also frequently observed in liposarcomas.  Although
such tumors retain the normal p53 gene, the increased levels of Mdm2 keep p53 in an
inactive state, hence, restricting their tumor suppression activity.

Cells Coordinate Growth and Proliferation

Overview

Cell size is a significant factor impacting cellular design, function, and fitness. There exists
some internal coordination by which cells double their masses before division, thus,
achieving homeostasis. Coordination between cell growth and proliferation depends on the
checkpoints in between cell cycle phases. Loss of coordination or failure in the checkpoint
mechanism can drive the cell to uncontrolled growth and loss of cellular function. Like
dividing cells that coordinate cellular growth, non-dividing cells in adults regulate cell size
depending on their metabolic states.

In adults, the size of non-dividing muscle cells varies depending on the environmental
conditions and nutritional state. Regular physical workout causes adult muscle cells to
enlarge as individual myocytes grow in size due to the absence of proliferation in the
myocytes themselves or the muscle stem cell population. In contrast, nutrient deficiency
can severely damage the muscle cells. The size of muscle cells depends on the balance
between the anabolic pathway that increases the cell size and the catabolic pathway that
degrades intracellular proteins causing the cell size to reduce.

The anabolic or IFF/PI3K/AKT/mTORC1 pathway involves mTORC1 signaling that leads to

protein synthesis, giving rise to a condition called- muscle cell hypertrophy. However, this
increase in cell size is temporary. For the cells to maintain their size, one must regularly
exercise for continuous mTORC1 signaling. Lack of exercise, starvation, or certain muscle
disease triggers a catabolic pathway or Myostatin/SMAD2/3 for protein degradation. The
degradation of proteins mobilizes amino acids to other cells of the body, thus, reducing the
size of the skeletal muscle cells.

Procedure

Cell size is a crucial factor in most fundamental processes, such as nutrient transport.

In an abnormally large cell, nutrients have to move longer distances to spread throughout
the cell. As a result, nutrient diffusion becomes slow, causing the cell to die from nutrient
starvation. 

Therefore, a healthy cell regulates its growth at cell cycle checkpoints- usually at the G1/S
phase transition or the G2/M phase transition.

These checkpoints enable the cell to monitor its size and regulate the timing of cell division,
thereby ensuring the daughter cells have a consistent size- a phenomenon called size
homeostasis.

Unicellular organisms, such as yeast, are frequently used as model organisms to study size
homeostasis.
A budding yeast cell divides asymmetrically, producing a larger mother cell and a smaller
daughter cell.

The larger mother cell quickly grows to its critical size and passes the size checkpoint at
the G1/S phase transition.

In contrast, the smaller daughter cell has a large size gap to achieve and therefore spends
more time growing in the G1 phase.

At the early G1 phase, protein complexes SBF and MBF, the cell cycle-promoting
transcription factors are usually bound and inhibited by a repressor protein called Whi5,
thereby preventing cell cycle transition.

The cell cycle reactivation depends on a sizer protein called Cln3, a G1 cyclin whose
concentration increases proportionally with the cell size.

When the cell attains its target size, Cln3 reaches a critical concentration and forms a
complex with cyclin-dependent kinase-1 or CDK1, a key activator of cell cycle promoting
factors. 

The active Cln3-CDK1 complex then phosphorylates Whi5 at multiple sites to release
active SBF and MBF transcription factors that trigger G1/S phase transition genes involved
in vital processes such as bud initiation and DNA replication.

Activation of these transition events enables the cell to pass the size checkpoint and
proceed through other stages of the cell cycle.