Chapter 10 | Cell Reproduction 279

10 | CELL
REPRODUCTION

Figure 10.1 A sea urchin begins life as a single diploid cell (zygote) that (a) divides through cell division to form two
genetically identical daughter cells, visible here through scanning electron microscopy (SEM). After four rounds of cell
division, (b) there are 16 cells, as seen in this SEM image. After many rounds of cell division, the individual develops
into a complex, multicellular organism, as seen in this (c) mature sea urchin. (credit a: modification of work by Evelyn
Spiegel, Louisa Howard; credit b: modification of work by Evelyn Spiegel, Louisa Howard; credit c: modification of work
by Marco Busdraghi; scale-bar data from Matt Russell)

Chapter Outline
10.1: Cell Division
10.2: The Cell Cycle
10.3: Control of the Cell Cycle
10.4: Cancer and the Cell Cycle
10.5: Prokaryotic Cell Division

Introduction
A human, like every sexually reproducing organism, begins life as a fertilized egg (embryo) or zygote. In our
species, billions of cell divisions subsequently must occur in a controlled manner in order to produce a complex,
multicellular human comprising trillions of cells. Thus, the original single-celled zygote is literally the ancestor
of all cells in the body. However, once a human is fully grown, cell reproduction is still necessary to repair and
regenerate tissues, and sometimes to increase our size! In fact, all multicellular organisms use cell division for
growth and the maintenance and repair of cells and tissues. Cell division is closely regulated, and the occasional
failure of this regulation can have life-threatening consequences. Single-celled organisms may also use cell
division as their method of reproduction.

10.1 | Cell Division

By the end of this section, you will be able to do the following:
• Describe the structure of prokaryotic and eukaryotic genomes
• Distinguish between chromosomes, genes, and traits
• Describe the mechanisms of chromosome compaction

The continuity of life from one cell to another has its foundation in the reproduction of cells by way of the cell
cycle. The cell cycle is an orderly sequence of events that describes the stages of a cell’s life from the division
of a single parent cell to the production of two new genetically identical daughter cells.
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Genomic DNA
Before discussing the steps a cell must undertake to replicate and divide its DNA, a deeper understanding of the
structure and function of a cell’s genetic information is necessary. A cell’s DNA, packaged as a double-stranded
DNA molecule, is called its genome. In prokaryotes, the genome is composed of a single, double-stranded DNA
molecule in the form of a loop or circle (Figure 10.2). The region in the cell containing this genetic material is
called a nucleoid. Some prokaryotes also have smaller loops of DNA called plasmids that are not essential for
normal growth. Bacteria can exchange these plasmids with other bacteria, sometimes receiving beneficial new
genes that the recipient can add to their chromosomal DNA. Antibiotic resistance is one trait that often spreads
through a bacterial colony through plasmid exchange from resistant donors to recipient cells.

Figure 10.2 Prokaryotes, including both Bacteria and Archaea, have a single, circular chromosome located in a central
region called the nucleoid.

In eukaryotes, the genome consists of several double-stranded linear DNA molecules (Figure 10.3). Each
species of eukaryotes has a characteristic number of chromosomes in the nuclei of its cells. Human body
(somatic) cells have 46 chromosomes, while human gametes (sperm or eggs) have 23 chromosomes each.
A typical body cell contains two matched or homologous sets of chromosomes (one set from each biological
parent)—a configuration known as diploid. (Note: The letter n is used to represent a single set of chromosomes;
therefore, a diploid organism is designated 2n.) Human cells that contain one set of chromosomes are called
gametes, or sex cells; these are eggs and sperm, and are designated 1n, or haploid.
Upon fertilization, each gamete contributes one set of chromosomes, creating a diploid cell containing matched
pairs of chromosomes called homologous (“same knowledge”) chromosomes. Homologous chromosomes are
the same length and have specific nucleotide segments called genes in exactly the same location, or locus.
Genes, the functional units of chromosomes, determine specific characteristics by coding for specific proteins.
Traits are the variations of those characteristics. For example, hair color is a characteristic with traits that are
blonde, brown, or black, and many colors in between.

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Chapter 10 | Cell Reproduction 281

Figure 10.3 There are 23 pairs of homologous chromosomes in a female human somatic cell. The condensed
chromosomes are viewed within the nucleus (top), removed from a cell during mitosis (also called karyokinesis or
nuclear division) and spread out on a slide (right), and artificially arranged according to length (left); an arrangement
like this is called a karyotype. In this image, the chromosomes were exposed to fluorescent stains for differentiation of
the different chromosomes. A method of staining called “chromosome painting” employs fluorescent dyes that highlight
chromosomes in different colors. (credit: National Human Genome Project/NIH)

Each copy of a homologous pair of chromosomes originates from a different parent; therefore, the different
genes (alleles) themselves are not identical, although they code for the same traits such as “hair color.” The
variation of individuals within a species is due to the specific combination of the genes inherited from both
parents. Even a slightly altered sequence of nucleotides within a gene can result in an alternative trait. For
example, there are three possible gene sequences on the human chromosome that code for blood type:
sequence A, sequence B, and sequence O. Because all diploid human cells have two copies of the chromosome
that determines blood type, the blood type (the trait) is determined by the two alleles of the marker gene that are
inherited. It is possible to have two copies of the same gene sequence on both homologous chromosomes, with
one on each (for example, AA, BB, or OO), or two different sequences, such as AB, AO, or BO.
Apparently minor variations of traits, such as blood type, eye color, and handedness, contribute to the natural
variation found within a species, but even though they seem minor, these traits may be connected with the
expression of other traits as of yet unknown. However, if the entire DNA sequence from any pair of human
homologous chromosomes is compared, the difference is much less than one percent. The sex chromosomes,
X and Y, are the single exception to the rule of homologous chromosome uniformity: Other than a small amount
of homology that is necessary to accurately produce gametes, the genes found on the X and Y chromosomes
are different.

Eukaryotic Chromosomal Structure and Compaction

If the DNA from all 46 chromosomes in a human cell nucleus were laid out end-to-end, it would measure
approximately two meters; however, its diameter would be only 2 nm! Considering that the size of a typical
human cell is about 10 µm (100,000 cells lined up to equal one meter), DNA must be tightly packaged to fit in
the cell’s nucleus. At the same time, it must also be readily accessible for the genes to be expressed. For this
reason, the long strands of DNA are condensed into compact chromosomes during certain stages of the cell
cycle. There are a number of ways that chromosomes are compacted.
In the first level of compaction, short stretches of the DNA double helix wrap around a core of eight histone
proteins at regular intervals along the entire length of the chromosome (Figure 10.4). The DNA-histone complex
is called chromatin. The beadlike, histone DNA complex is called a nucleosome, and DNA connecting the
nucleosomes is called linker DNA. A DNA molecule in this form is about seven times shorter than the double
helix without the histones, and the beads are about 10 nm in diameter, in contrast with the 2-nm diameter of a
DNA double helix.
The second level of compaction occurs as the nucleosomes and the linker DNA between them coil into a 30-nm
chromatin fiber. This coiling further condenses the chromosome so that it is now about 50 times shorter than the
extended form.
282 Chapter 10 | Cell Reproduction

In the third level of compaction, a variety of fibrous proteins is used to “pack the chromatin.” These fibrous
proteins also ensure that each chromosome in a non-dividing cell occupies a particular area of the nucleus that
does not overlap with that of any other chromosome (see the top image in Figure 10.3).

Figure 10.4 Double-stranded DNA wraps around histone proteins to form nucleosomes that create the appearance
of “beads on a string.” The nucleosomes are coiled into a 30-nm chromatin fiber. When a cell undergoes mitosis, the
chromosomes condense even further.

DNA replicates in the S phase of interphase, which technically is not a part of mitosis, but must always precede it.
After replication, the chromosomes are composed of two linked sister chromatids. When fully compact, the pairs
of identically packed chromosomes are bound to each other by cohesin proteins. The connection between the
sister chromatids is closest in a region called the centromere. The conjoined sister chromatids, with a diameter
of about 1 µm, are visible under a light microscope. The centromeric region is highly condensed and thus will
appear as a constricted area.

This animation illustrates the different levels of chromosome packing.

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id1165802978636)

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Chapter 10 | Cell Reproduction 283

10.2 | The Cell Cycle

By the end of this section, you will be able to do the following:
• Describe the three stages of interphase
• Discuss the behavior of chromosomes during karyokinesis/mitosis
• Explain how the cytoplasmic content is divided during cytokinesis
• Define the quiescent G0 phase

The cell cycle is an ordered series of events involving cell growth and cell division that produces two new
daughter cells. Cells on the path to cell division proceed through a series of precisely timed and carefully
regulated stages of growth, DNA replication, and nuclear and cytoplasmic division that ultimately produces two
identical (clone) cells. The cell cycle has two major phases: interphase and the mitotic phase (Figure 10.5).
During interphase, the cell grows and DNA is replicated. During the mitotic phase, the replicated DNA and
cytoplasmic contents are separated, and the cell cytoplasm is typically partitioned by a third process of the cell
cycle called cytokinesis. We should note, however, that interphase and mitosis (kayrokinesis) may take place
without cytokinesis, in which case cells with multiple nuclei (multinucleate cells) are produced.

Figure 10.5 The cell cycle in multicellular organisms consists of interphase and the mitotic phase. During interphase,
the cell grows and the nuclear DNA is duplicated. Interphase is followed by the mitotic phase. During the mitotic phase,
the duplicated chromosomes are segregated and distributed into daughter nuclei. Following mitosis, the cytoplasm is
usually divided as well by cytokinesis, resulting in two genetically identical daughter cells.

Interphase
During interphase, the cell undergoes normal growth processes while also preparing for cell division. In order for
a cell to move from interphase into the mitotic phase, many internal and external conditions must be met. The
three stages of interphase are called G1, S, and G2.
G1 Phase (First Gap)
The first stage of interphase is called the G1 phase (first gap) because, from a microscopic point of view, little
284 Chapter 10 | Cell Reproduction

change is visible. However, during the G1 stage, the cell is quite active at the biochemical level. The cell is
accumulating the building blocks of chromosomal DNA and the associated proteins as well as accumulating
sufficient energy reserves to complete the task of replicating each chromosome in the nucleus.
S Phase (Synthesis of DNA)
Throughout interphase, nuclear DNA remains in a semi-condensed chromatin configuration. In the S phase,
DNA replication can proceed through the mechanisms that result in the formation of identical pairs of DNA
molecules—sister chromatids—that are firmly attached to the centromeric region. The centrosome is also
duplicated during the S phase. The two centrosomes of homologous chromosomes will give rise to the mitotic
spindle, the apparatus that orchestrates the movement of chromosomes during mitosis. For example, roughly
at the center of each animal cell, the centrosomes are associated with a pair of rod-like objects, the centrioles,
which are positioned at right angles to each other. Centrioles help organize cell division. We should note,
however, that centrioles are not present in the centrosomes of other eukaryotic organisms, such as plants and
most fungi.
G2 Phase (Second Gap)
In the G2 phase, the cell replenishes its energy stores and synthesizes proteins necessary for chromosome
manipulation and movement. Some cell organelles are duplicated, and the cytoskeleton is dismantled to provide
resources for the mitotic phase. There may be additional cell growth during G2. The final preparations for the
mitotic phase must be completed before the cell is able to enter the first stage of mitosis.

The Mitotic Phase

The mitotic phase is a multistep process during which the duplicated chromosomes are aligned, separated,
and move into two new, identical daughter cells. The first portion of the mitotic phase is called karyokinesis,
or nuclear division. As we have just seen, the second portion of the mitotic phase (and often viewed as a
process separate from and following mitosis) is called cytokinesis—the physical separation of the cytoplasmic
components into the two daughter cells.

Revisit the stages of mitosis at this site (http://openstaxcollege.org/l/Cell_cycle_mito) .

Karyokinesis (Mitosis)
Karyokinesis, also known as mitosis, is divided into a series of phases—prophase, prometaphase, metaphase,
anaphase, and telophase—that result in the division of the cell nucleus (Figure 10.6).

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Chapter 10 | Cell Reproduction 285

Figure 10.6 Karyokinesis (or mitosis) is divided into five stages—prophase, prometaphase, metaphase, anaphase,
and telophase. We should note that this is a continuous process, and that the divisions between the stages are
not discrete. The pictures at the bottom were taken by fluorescence microscopy (hence, the black background) of
cells artificially stained by fluorescent dyes: blue fluorescence indicates DNA (chromosomes) and green fluorescence
indicates microtubules (spindle apparatus). (credit “mitosis drawings”: modification of work by Mariana Ruiz Villareal;
credit “micrographs”: modification of work by Roy van Heesbeen; credit “cytokinesis micrograph”: Wadsworth Center/
New York State Department of Health; scale-bar data from Matt Russell)

Prophase (the “first phase”): the nuclear envelope starts to dissociate into small vesicles, and the membranous
organelles (such as the Golgi complex [Golgi apparatus] and the endoplasmic reticulum), fragment and disperse
toward the periphery of the cell. The nucleolus disappears (disperses) as well, and the centrosomes begin
to move to opposite poles of the cell. Microtubules that will form the mitotic spindle extend between the
centrosomes, pushing them farther apart as the microtubule fibers lengthen. The sister chromatids begin to coil
more tightly with the aid of condensin proteins and now become visible under a light microscope.
Prometaphase (the “first change phase”): Many processes that began in prophase continue to advance. The
remnants of the nuclear envelope fragment further, and the mitotic spindle continues to develop as more
microtubules assemble and stretch across the length of the former nuclear area. Chromosomes become even
more condensed and discrete. Each sister chromatid develops a protein structure called a kinetochore in
its centromeric region (Figure 10.7). The proteins of the kinetochore attract and bind to the mitotic spindle
microtubules. As the spindle microtubules extend from the centrosomes, some of these microtubules come into
contact with and firmly bind to the kinetochores. Once a mitotic fiber attaches to a chromosome, the chromosome
will be oriented until the kinetochores of sister chromatids face the opposite poles. Eventually, all the sister
chromatids will be attached via their kinetochores to microtubules from opposing poles. Spindle microtubules
that do not engage the chromosomes are called polar microtubules. These microtubules overlap each other
midway between the two poles and contribute to cell elongation. Astral microtubules are located near the poles,
aid in spindle orientation, and are required for the regulation of mitosis.
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Figure 10.7 During prometaphase, mitotic spindle microtubules from opposite poles attach to each sister chromatid at
the kinetochore. In anaphase, the connection between the sister chromatids breaks down, and the microtubules pull
the chromosomes toward opposite poles.

Metaphase (the “change phase”): All the chromosomes are aligned in a plane called the metaphase plate, or
the equatorial plane, roughly midway between the two poles of the cell. The sister chromatids are still tightly
attached to each other by cohesin proteins. At this time, the chromosomes are maximally condensed.
Anaphase (“upward phase”): The cohesin proteins degrade, and the sister chromatids separate at the
centromere. Each chromatid, now called a single chromosome, is pulled rapidly toward the centrosome to which
its microtubule is attached. The cell becomes visibly elongated (oval shaped) as the polar microtubules slide
against each other at the metaphase plate where they overlap.
Telophase (the “distance phase”): the chromosomes reach the opposite poles and begin to decondense
(unravel), relaxing once again into a stretched-out chromatin configuration. The mitotic spindles are
depolymerized into tubulin monomers that will be used to assemble cytoskeletal components for each daughter
cell. Nuclear envelopes form around the chromosomes, and nucleosomes appear within the nuclear area.
Cytokinesis
Cytokinesis, or “cell motion,” is sometimes viewed as the second main stage of the mitotic phase, during which
cell division is completed via the physical separation of the cytoplasmic components into two daughter cells
However, as we have seen earlier, cytokinesis can also be viewed as a separate phase, which may or may not
take place following mitosis. If cytokinesis does take place, cell division is not complete until the cell components
have been apportioned and completely separated into the two daughter cells. Although the stages of mitosis are
similar for most eukaryotes, the process of cytokinesis is quite different for eukaryotes that have cell walls, such
as plant cells.
In animal cells, cytokinesis typically starts during late anaphase. A contractile ring composed of actin filaments
forms just inside the plasma membrane at the former metaphase plate. The actin filaments pull the equator of
the cell inward, forming a fissure. This fissure is called the cleavage furrow. The furrow deepens as the actin
ring contracts, and eventually the membrane is cleaved in two (Figure 10.8).
In plant cells, a new cell wall must form between the daughter cells. During interphase, the Golgi apparatus
accumulates enzymes, structural proteins, and glucose molecules prior to breaking into vesicles and dispersing
throughout the dividing cell. During telophase, these Golgi vesicles are transported on microtubules to form a
phragmoplast (a vesicular structure) at the metaphase plate. There, the vesicles fuse and coalesce from the
center toward the cell walls; this structure is called a cell plate. As more vesicles fuse, the cell plate enlarges
until it merges with the cell walls at the periphery of the cell. Enzymes use the glucose that has accumulated
between the membrane layers to build a new cell wall. The Golgi membranes become parts of the plasma
membrane on either side of the new cell wall (Figure 10.8).

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Chapter 10 | Cell Reproduction 287

Figure 10.8 During cytokinesis in animal cells, a ring of actin filaments forms at the metaphase plate. The ring
contracts, forming a cleavage furrow, which divides the cell in two. In plant cells, Golgi vesicles coalesce at the former
metaphase plate, forming a phragmoplast. A cell plate formed by the fusion of the vesicles of the phragmoplast grows
from the center toward the cell walls, and the membranes of the vesicles fuse to form a plasma membrane that divides
the cell in two.

G0 Phase
Not all cells adhere to the classic cell-cycle pattern in which a newly formed daughter cell immediately enters
the preparatory phases of interphase, closely followed by the mitotic phase, and cytokinesis. Cells in G0 phase
are not actively preparing to divide. The cell is in a quiescent (inactive) stage that occurs when cells exit the
cell cycle. Some cells enter G0 temporarily due to environmental conditions such as availability of nutrients, or
stimulation by growth factors. The cell will remain in this phase until conditions improve or until an external signal
triggers the onset of G1. Other cells that never or rarely divide, such as mature cardiac muscle and nerve cells,
remain in G0 permanently.

Which of the following is the correct order of events in mitosis?

a. Sister chromatids line up at the metaphase plate. The kinetochore becomes attached to the mitotic
spindle. The nucleus reforms and the cell divides. Cohesin proteins break down and the sister
chromatids separate.
b. The kinetochore becomes attached to the mitotic spindle. Cohesin proteins break down and the sister
chromatids separate. Sister chromatids line up at the metaphase plate. The nucleus reforms and the
cell divides.
c. The kinetochore becomes attached to the cohesin proteins. Sister chromatids line up at the metaphase
plate. The kinetochore breaks down and the sister chromatids separate. The nucleus reforms and the
cell divides.
d. The kinetochore becomes attached to the mitotic spindle. Sister chromatids line up at the metaphase
plate. Cohesin proteins break down and the sister chromatids separate. The nucleus reforms and the
cell divides.
288 Chapter 10 | Cell Reproduction

Determine the Time Spent in Cell-Cycle Stages

Problem: How long does a cell spend in interphase compared to each stage of mitosis?
Background: A prepared microscope slide of whitefish blastula cross-sections will show cells arrested in
various stages of the cell cycle. (Note: It is not visually possible to separate the stages of interphase from
each other, but the mitotic stages are readily identifiable.) If 100 cells are examined, the number of cells in
each identifiable cell-cycle stage will give an estimate of the time it takes for the cell to complete that stage.
Problem Statement: Given the events included in all of interphase and those that take place in each stage
of mitosis, estimate the length of each stage based on a 24-hour cell cycle. Before proceeding, state your
hypothesis.
Test your hypothesis: Test your hypothesis by doing the following:
1. Place a fixed and stained microscope slide of whitefish blastula cross-sections under the scanning
objective of a light microscope.
2. Locate and focus on one of the sections using the low-power objective of your microscope. Notice that
the section is a circle composed of dozens of closely packed individual cells.
3. Switch to the medium-power objective and refocus. With this objective, individual cells are clearly
visible, but the chromosomes will still be very small.
4. Switch to the high-power objective and slowly move the slide left to right, and up and down to view
all the cells in the section (Figure 10.9). As you scan, you will notice that most of the cells are not
undergoing mitosis but are in the interphase period of the cell cycle.

(b)

(a)
Figure 10.9 Slowly scan whitefish blastula cells with the high-power objective as illustrated in image (a)
to identify their mitotic stage. (b) A microscopic image of the scanned cells is shown. (credit “micrograph”:
modification of work by Linda Flora; scale-bar data from Matt Russell)

5. Practice identifying the various stages of the cell cycle, using the drawings of the stages as a guide
(Figure 10.6).
6. Once you are confident about your identification, begin to record the stage of each cell you encounter
as you scan left to right, and top to bottom across the blastula section.
7. Keep a tally of your observations and stop when you reach 100 cells identified.
8. The larger the sample size (total number of cells counted), the more accurate the results. If possible,
gather and record group data prior to calculating percentages and making estimates.
Record your observations: Make a table similar to Table 10.1 within which to record your observations.

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Chapter 10 | Cell Reproduction 289

Results of Cell Stage Identification

Phase or Stage Individual Totals Group Totals Percent
Interphase
Prophase
Metaphase
Anaphase
Telophase
Cytokinesis
Totals 100 100 100 percent

Table 10.1

Analyze your data/report your results: To find the length of time whitefish blastula cells spend in each
stage, multiply the percent (recorded as a decimal) by 24 hours. Make a table similar to Table 10.2 to
illustrate your data.

Estimate of Cell Stage Length

Phase or Stage Percent Time in Hours
Interphase
Prophase
Metaphase
Anaphase
Telophase
Cytokinesis

Table 10.2

Draw a conclusion: Did your results support your estimated times? Were any of the outcomes unexpected?
If so, discuss those events in that stage that may have contributed to the calculated time.

10.3 | Control of the Cell Cycle

By the end of this section, you will be able to do the following:
• Understand how the cell cycle is controlled by mechanisms that are both internal and external to the cell
• Explain how the three internal “control checkpoints” occur at the end of G1, at the G2/M transition, and
during metaphase
• Describe the molecules that control the cell cycle through positive and negative regulation

The length of the cell cycle is highly variable, even within the cells of a single organism. In humans, the frequency
of cell turnover ranges from a few hours in early embryonic development, to an average of two to five days for
epithelial cells, and to an entire human lifetime spent in G0 by specialized cells, such as cortical neurons or
cardiac muscle cells.
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There is also variation in the time that a cell spends in each phase of the cell cycle. When rapidly dividing
mammalian cells are grown in a culture (outside the body under optimal growing conditions), the length of
the cell cycle is about 24 hours. In rapidly dividing human cells with a 24-hour cell cycle, the G1 phase lasts
approximately nine hours, the S phase lasts 10 hours, the G2 phase lasts about four and one-half hours, and the
M phase lasts approximately one-half hour. By comparison, in fertilized eggs (and early embryos) of fruit flies,
the cell cycle is completed in about eight minutes. This is because the nucleus of the fertilized egg divides many
times by mitosis but does not go through cytokinesis until a multinucleate “zygote” has been produced, with
many nuclei located along the periphery of the cell membrane, thereby shortening the time of the cell division
cycle. The timing of events in the cell cycle of both “invertebrates” and “vertebrates” is controlled by mechanisms
that are both internal and external to the cell.

Regulation of the Cell Cycle by External Events

Both the initiation and inhibition of cell division are triggered by events external to the cell when it is about to
begin the replication process. An event may be as simple as the death of nearby cells or as sweeping as the
release of growth-promoting hormones, such as human growth hormone (HGH or hGH). A lack of HGH can
inhibit cell division, resulting in dwarfism, whereas too much HGH can result in gigantism. Crowding of cells can
also inhibit cell division. In contrast, a factor that can initiate cell division is the size of the cell: As a cell grows, it
becomes physiologically inefficient due to its decreasing surface-to-volume ratio. The solution to this problem is
to divide.
Whatever the source of the message, the cell receives the signal, and a series of events within the cell allows it
to proceed into interphase. Moving forward from this initiation point, every parameter required during each cell
cycle phase must be met or the cycle cannot progress.

Regulation at Internal Checkpoints

It is essential that the daughter cells produced be exact duplicates of the parent cell. Mistakes in the duplication
or distribution of the chromosomes lead to mutations that may be passed forward to every new cell produced
from an abnormal cell. To prevent a compromised cell from continuing to divide, there are internal control
mechanisms that operate at three main cell-cycle checkpoints: A checkpoint is one of several points in the
eukaryotic cell cycle at which the progression of a cell to the next stage in the cycle can be halted until conditions
are favorable. These checkpoints occur near the end of G1, at the G2/M transition, and during metaphase
(Figure 10.10).

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Chapter 10 | Cell Reproduction 291

Figure 10.10 The cell cycle is controlled at three checkpoints. The integrity of the DNA is assessed at the G1
checkpoint. Proper chromosome duplication is assessed at the G2 checkpoint. Attachment of each kinetochore to a
spindle fiber is assessed at the M checkpoint.

The G1 Checkpoint
The G1 checkpoint determines whether all conditions are favorable for cell division to proceed. The G1
checkpoint, also called the restriction point (in yeast), is a point at which the cell irreversibly commits to the cell
division process. External influences, such as growth factors, play a large role in carrying the cell past the G1
checkpoint. In addition to adequate reserves and cell size, there is a check for genomic DNA damage at the
G1 checkpoint. A cell that does not meet all the requirements will not be allowed to progress into the S phase.
The cell can halt the cycle and attempt to remedy the problematic condition, or the cell can advance into G0 and
await further signals when conditions improve.
The G2 Checkpoint
The G2 checkpoint bars entry into the mitotic phase if certain conditions are not met. As at the G1 checkpoint,
cell size and protein reserves are assessed. However, the most important role of the G2 checkpoint is to ensure
that all of the chromosomes have been replicated and that the replicated DNA is not damaged. If the checkpoint
mechanisms detect problems with the DNA, the cell cycle is halted, and the cell attempts to either complete DNA
replication or repair the damaged DNA.
The M Checkpoint
The M checkpoint occurs near the end of the metaphase stage of karyokinesis. The M checkpoint is also known
as the spindle checkpoint, because it determines whether all the sister chromatids are correctly attached to the
spindle microtubules. Because the separation of the sister chromatids during anaphase is an irreversible step,
the cycle will not proceed until the kinetochores of each pair of sister chromatids are firmly anchored to at least
two spindle fibers arising from opposite poles of the cell.
292 Chapter 10 | Cell Reproduction

Watch what occurs at the G1, G2, and M checkpoints by visiting this website (http://openstaxcollege.org/l/
cell_checkpnts) to see an animation of the cell cycle.

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. However, 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 termed positive regulators.
They 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 (Figure 10.11). 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 by cytoplasmic enzymes, as shown in
Figure 10.11 below.

Figure 10.11 The concentrations of cyclin proteins change throughout the cell cycle. There is a direct correlation
between cyclin accumulation and the three major cell-cycle checkpoints. Also note the sharp decline of cyclin levels
following each checkpoint (the transition between phases of the cell cycle), as cyclin is degraded by cytoplasmic
enzymes. (credit: modification of work by "WikiMiMa"/Wikimedia Commons)

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 to activate the complex. Like all kinases, Cdks are enzymes
(kinases) that in turn 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. (Figure 10.12).
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.

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Chapter 10 | Cell Reproduction 293

Figure 10.12 Cyclin-dependent kinases (Cdks) are protein kinases that, when fully activated, can phosphorylate and
thus activate other proteins that advance the cell cycle past a checkpoint. To become fully activated, a Cdk must bind
to a cyclin protein and then be phosphorylated by another kinase.

Because the cyclic fluctuations of cyclin levels are largely based on the timing of the cell cycle and not on specific
events, regulation of the cell cycle usually occurs by either the Cdk molecules alone or the Cdk/cyclin complexes.
Without a specific concentration of fully activated cyclin/Cdk complexes, the cell cycle cannot proceed through
the 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 that the inhibitor monitors is completed.
294 Chapter 10 | Cell Reproduction

Negative Regulation of the Cell Cycle

The second group of cell-cycle regulatory molecules are negative regulators, which stop 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. We should note here
that the 53 and 21 designations refer to the functional molecular masses of the proteins (p) in kilodaltons (a
dalton is equal to an atomic mass unit, which is equal to one proton or one neutron or 1 g/mol). 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 (i.e., 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 commitment of a cell to division because 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 then recruits specific
enzymes to repair the DNA. If the DNA cannot be repaired, p53 can trigger apoptosis, or 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, which largely monitors cell size, exerts its regulatory influence on other positive regulator proteins. In the
active, dephosphorylated state, Rb binds to proteins called transcription factors, most commonly, E2F (Figure
10.13). 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.”

Figure 10.13 Rb halts the cell cycle and releases its hold in response to cell growth.

Rb and other proteins that negatively regulate the cell cycle are sometimes called tumor suppressors. Why
do you think the name tumor suppressor might be appropriate for these proteins?

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Chapter 10 | Cell Reproduction 295

10.4 | Cancer and the Cell Cycle

By the end of this section, you will be able to do the following:
• Describe how cancer is caused by uncontrolled cell growth
• Understand how proto-oncogenes are normal cell genes that, when mutated, become oncogenes
• Describe how tumor suppressors function
• Explain how mutant tumor suppressors cause cancer

Cancer comprises many different diseases caused by a common mechanism: uncontrolled cell growth. Despite
the redundancy and overlapping levels of cell-cycle control, errors do occur. One of the critical processes
monitored by the cell-cycle checkpoint surveillance mechanism is the proper replication of DNA during the S
phase. Even when all of the cell-cycle controls are fully functional, a small percentage of replication errors
(mutations) will be passed on to the daughter cells. If changes to the DNA nucleotide sequence occur within a
coding portion of a gene and are not corrected, a gene mutation results. All cancers start when a gene mutation
gives rise to a faulty protein that plays a key role in cell reproduction.
The change in the cell that results from the malformed protein may be minor: perhaps a slight delay in the
binding of Cdk to cyclin or an Rb protein that detaches from its target DNA while still phosphorylated. Even minor
mistakes, however, may allow subsequent mistakes to occur more readily. Over and over, small uncorrected
errors are passed from the parent cell to the daughter cells and amplified as each generation produces more
non-functional proteins from uncorrected DNA damage. Eventually, the pace of the cell cycle speeds up as
the effectiveness of the control and repair mechanisms decreases. Uncontrolled growth of the mutated cells
outpaces the growth of normal cells in the area, and a tumor (“-oma”) can result.

Proto-oncogenes
The genes that code for the positive cell-cycle regulators are called proto-oncogenes. Proto-oncogenes are
normal genes that, when mutated in certain ways, become oncogenes—genes that cause a cell to become
cancerous. Consider what might happen to the cell cycle in a cell with a recently acquired oncogene. In most
instances, the alteration of the DNA sequence will result in a less functional (or non-functional) protein. The result
is detrimental to the cell and will likely prevent the cell from completing the cell cycle; however, the organism
is not harmed because the mutation will not be carried forward. If a cell cannot reproduce, the mutation is
not propagated and the damage is minimal. Occasionally, however, a gene mutation causes a change that
increases the activity of a positive regulator. For example, a mutation that allows Cdk to be activated without
being partnered with cyclin could push the cell cycle past a checkpoint before all of the required conditions are
met. If the resulting daughter cells are too damaged to undergo further cell divisions, the mutation would not be
propagated and no harm would come to the organism. However, if the atypical daughter cells are able to undergo
further cell divisions, subsequent generations of cells may accumulate even more mutations, some possibly in
additional genes that regulate the cell cycle.
The Cdk gene in the above example is only one of many genes that are considered proto-oncogenes. In addition
to the cell-cycle regulatory proteins, any protein that influences the cycle can be altered in such a way as to
override cell-cycle checkpoints. An oncogene is any gene that, when altered, leads to an increase in the rate of
cell-cycle progression.

Tumor Suppressor Genes

Like proto-oncogenes, many of the negative cell-cycle regulatory proteins were discovered in cells that had
become cancerous. Tumor suppressor genes are segments of DNA that code for negative regulator proteins,
the type of regulators that, when activated, can prevent the cell from undergoing uncontrolled division. The
collective function of the best-understood tumor suppressor gene proteins, Rb, p53, and p21, is to put up a
roadblock to cell-cycle progression until certain events are completed. A cell that carries a mutated form of a
negative regulator might not be able to halt the cell cycle if there is a problem. Tumor suppressors are similar to
brakes in a vehicle: Malfunctioning brakes can contribute to a car crash!
Mutated p53 genes have been identified in more than 50 percent of all human tumor cells. This discovery is not
surprising in light of the multiple roles that the p53 protein plays at the G1 checkpoint. A cell with a faulty p53 may
296 Chapter 10 | Cell Reproduction

fail to detect errors present in the genomic DNA (Figure 10.14). Even if a partially functional p53 does identify
the mutations, it may no longer be able to signal the necessary DNA repair enzymes. Either way, damaged DNA
will remain uncorrected. At this point, a functional p53 will deem the cell unsalvageable and trigger programmed
cell death (apoptosis). The damaged version of p53 found in cancer cells, however, cannot trigger apoptosis.

Figure 10.14 The role of normal p53 is to monitor DNA and the supply of oxygen (hypoxia is a condition of reduced
oxygen supply). If damage is detected, p53 triggers repair mechanisms. If repairs are unsuccessful, p53 signals
apoptosis. A cell with an abnormal p53 protein cannot repair damaged DNA and thus cannot signal apoptosis.
Cells with abnormal p53 can become cancerous. (credit: modification of work by Thierry Soussi)

Human papillomavirus can cause cervical cancer. The virus encodes E6, a protein that binds p53. Based
on this fact and what you know about p53, what effect do you think E6 binding has on p53 activity?
a. E6 activates p53
b. E6 inactivates p53
c. E6 mutates p53
d. E6 binding marks p53 for degradation

The loss of p53 function has other repercussions for the cell cycle. Mutated p53 might lose its ability to trigger
p21 production. Without adequate levels of p21, there is no effective block on Cdk activation. Essentially, without
a fully functional p53, the G1 checkpoint is severely compromised and the cell proceeds directly from G1 to
S regardless of internal and external conditions. At the completion of this shortened cell cycle, two daughter
cells are produced that have inherited the mutated p53 gene. Given the non-optimal conditions under which the
parent cell reproduced, it is likely that the daughter cells will have acquired other mutations in addition to the
faulty tumor-suppressor gene. Cells such as these daughter cells quickly accumulate both oncogenes and non-
functional tumor-suppressor genes. Again, the result is tumor growth.

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Chapter 10 | Cell Reproduction 297

Watch an animation of how cancer results from errors in the cell cycle. (This multimedia resource will open
in a browser.) (http://cnx.org/content/m66480/1.3/#eip-id1169995709332)

10.5 | Prokaryotic Cell Division

By the end of this section, you will be able to do the following:
• Describe the process of binary fission in prokaryotes
• Explain how FtsZ and tubulin proteins are examples of homology

Prokaryotes, such as bacteria, produce daughter cells by binary fission. For unicellular organisms, cell division
is the only method to produce new individuals. In both prokaryotic and eukaryotic cells, the outcome of cell
reproduction is a pair of daughter cells that are genetically identical to the parent cell. In unicellular organisms,
daughter cells are individuals.
To achieve the outcome of cloned offspring, certain steps are essential. The genomic DNA must be replicated
and then allocated into the daughter cells; the cytoplasmic contents must also be divided to give both new cells
the cellular machinery to sustain life. As we’ve seen with bacterial cells, the genome consists of a single, circular
DNA chromosome; therefore, the process of cell division is simplified. Karyokinesis is unnecessary because
there is no true nucleus and thus no need to direct one copy of the multiple chromosomes into each daughter
cell. This type of cell division is called binary (prokaryotic) fission.

Binary Fission
Due to the relative simplicity of the prokaryotes, the cell division process is a less complicated and much
more rapid process than cell division in eukaryotes. As a review of the general information on cell division
we discussed at the beginning of this chapter, recall that the single, circular DNA chromosome of bacteria
occupies a specific location, the nucleoid region, within the cell (Figure 10.2). Although the DNA of the nucleoid
is associated with proteins that aid in packaging the molecule into a compact size, there are no histone proteins
and thus no nucleosomes in prokaryotes. The packing proteins of bacteria are, however, related to the cohesin
and condensin proteins involved in the chromosome compaction of eukaryotes.
The bacterial chromosome is attached to the plasma membrane at about the midpoint of the cell. The starting
point of replication, the origin, is close to the binding site of the chromosome to the plasma membrane
(Figure 10.15). Replication of the DNA is bidirectional, moving away from the origin on both strands of the
loop simultaneously. As the new double strands are formed, each origin point moves away from the cell
wall attachment toward the opposite ends of the cell. As the cell elongates, the growing membrane aids in
the transport of the chromosomes. After the chromosomes have cleared the midpoint of the elongated cell,
cytoplasmic separation begins. The formation of a ring composed of repeating units of a protein called FtsZ
(short for “filamenting temperature-sensitive mutant Z”) directs the partition between the nucleoids. Formation of
the FtsZ ring triggers the accumulation of other proteins that work together to recruit new membrane and cell
wall materials to the site. A septum is formed between the daughter nucleoids, extending gradually from the
periphery toward the center of the cell. When the new cell walls are in place, the daughter cells separate.
298 Chapter 10 | Cell Reproduction

Figure 10.15 These images show the steps of binary fission in prokaryotes. (credit: modification of work by
“Mcstrother”/Wikimedia Commons)

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Chapter 10 | Cell Reproduction 299

Mitotic Spindle Apparatus

The precise timing and formation of the mitotic spindle is critical to the success of eukaryotic cell division.
Prokaryotic cells, on the other hand, do not undergo karyokinesis and therefore have no need for a mitotic
spindle. However, the FtsZ protein that plays such a vital role in prokaryotic cytokinesis is structurally
and functionally very similar to tubulin, the building block of the microtubules which make up the mitotic
spindle fibers that are necessary for eukaryotic nuclear division. FtsZ proteins can form filaments, rings,
and other three-dimensional structures that resemble the way tubulin forms microtubules, centrioles, and
various cytoskeletal components. In addition, both FtsZ and tubulin employ the same energy source, GTP
(guanosine triphosphate), to rapidly assemble and disassemble complex structures.
FtsZ and tubulin are considered to be homologous structures derived from common evolutionary origins. In
this example, FtsZ is the ancestor protein to tubulin (an evolutionarily derived protein). While both proteins
are found in extant organisms, tubulin function has evolved and diversified tremendously since evolving
from its FtsZ prokaryotic origin. A survey of mitotic assembly components found in present-day unicellular
eukaryotes reveals crucial intermediary steps to the complex membrane-enclosed genomes of multicellular
eukaryotes (Table 10.3).

Cell Division Apparatus among Various Organisms

Separation
Structure of of
Division of nuclear material
genetic material daughter
cells
There is no nucleus. FtsZ proteins
Occurs through binary fission. As the
The single, circular assemble into a
chromosome is replicated, the two copies move
Prokaryotes chromosome exists in a ring that
to opposite ends of the cell by an unknown
region of cytoplasm pinches the cell
mechanism.
called the nucleoid. in two.
Microfilaments
Chromosomes attach to the nuclear envelope, form a
Some Linear chromosomes which remains intact. The mitotic spindle passes cleavage
protists exist in the nucleus. through the envelope and elongates the cell. No furrow that
centrioles exist. pinches the cell
in two.
Microfilaments
A mitotic spindle forms from the centrioles and
Linear chromosomes form a
passes through the nuclear membrane, which
Other wrapped around cleavage
remains intact. Chromosomes attach to the
protists histones exist in the furrow that
mitotic spindle, which separates the
nucleus. pinches the cell
chromosomes and elongates the cell.
in two.
Microfilaments
A mitotic spindle forms from the centrosomes. form a
Linear chromosomes The nuclear envelope dissolves. Chromosomes cleavage
Animal cells
exist in the nucleus. attach to the mitotic spindle, which separates the furrow that
chromosomes and elongates the cell. pinches the cell
in two.

Table 10.3