Eukaryotic cell cycle

Elisa Giupponi, PhD student

Dipartimento di Chimica, Materiali e Ing. Chimica Giulio Natta

Politecnico Di Milano

Office: (+39)02-2399.4741
e-mail: elisa.giupponi@polimi.it
Outline

Cell cycle

Eukaryotic cell cycle

Mitosis and meiosis

Flow cytometry and cell cycle

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 Cell cycle

The division cycle of somatic eukaryotic cells

consists of four coordinated processes: cell
growth, DNA replication, distribution of the
duplicated chromosomes to daughter cells, and
cell division.

In bacteria, cell growth and DNA replication

take place throughout most of the cell cycle
and chromosomes are duplicated and
distributed to daughter cells in association with
plasma membrane in a process called binary
fission.

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Eukaryotic cell cycle
The eukaryotic cell cycle is divided into two
basic parts: M phase and interphase.
Mitosis and cytokinesis (M phase) last only
about an hour, so approximately 95% of the cell
cycle is spent in interphase. Cell cycle duration
varies according to the cell type, but for a
typical proliferating human cell the cycle lasts
about 24 h.

The interphase is divided in:

1. G1 phase
2. S phase
3. G2 phase
Progression between the stages of the cell cycle is
controlled by a regulatory apparatus, which not only
M phase is divided in: coordinates the different events of the cell cycle but
1. Mitosis also those generated by extracellular signals (e.g.,
growth factors) that control cell proliferation.
2. Cytokinesis

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Eukaryotic cell cycle
A)

During interphase, the chromosomes are

decondensed and distributed throughout
the nucleus.

G1 phase (gap 1): corresponds to the

interval between mitosis (M phase) and
initiation of DNA replication (S phase). Each
chromosome consists of 1 chromatid. B)
During G1 the cell is metabolically active and
continuously grows but does not replicate
its DNA. At a certain point (restriction point)
the cell moves to S phase.
S phase (DNA synthesis): DNA replication
takes place. Each chromosome consists of
two sister chromatids.
G2 phase (gap 2): cell growth continues
and proteins are synthesized in preparation
for mitosis. The cytoplasmic material is
prepared for mitosis and cytokinesis. A) Phases of the cell cycle

B) DNA doubling and chromosomal segregation during the cell

cycle

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Eukaryotic cell cycle
The eukaryotic cell cycle is divided into two
basic parts: M phase and interphase.
Mitosis and cytokinesis (M phase) last only
about an hour, so approximately 95% of the cell
cycle is spent in interphase. Cell cycle duration
varies according to the cell type, but for a
typical proliferating human cell the cycle lasts
about 24 h.

The interphase is divided in:

1. G1 phase
2. S phase
3. G2 phase
Progression between the stages of the cell cycle is
controlled by a regulatory apparatus, which not only
M phase is divided in: coordinates the different events of the cell cycle but
1. Mitosis also those generated by extracellular signals (e.g.,
growth factors) that control cell proliferation.
2. Cytokinesis

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Regulation of cell cycle: checkpoints
It is critically important that cells do not begin mitosis until the DNA duplication is completed, in
order to avoid that daughter cells inherit only a part of the genetic material.

Cells have feedback controls that prevent entry into the next phase until the preceding phase has
been completed.

There are 3 checkpoints that prevent the cell from duplicate damaged DNA or distribute an
incomplete part of the genome to the daughter cells.

G1 checkpoint arrests cell cycle and allows to repair the DNA

damage (e.g., effect of irradiation) to avoid the amplification of
the damage in the daughter cells.
G2 checkpoint prevents the initiation of mitosis if DNA is not fully
replicated or if it is damaged.
M checkpoint controls the alignment of chromosomes on the
mitotic spindle, ensuring that a complete set of chromosomes is
distributed to each of the daughter cells.

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Definitions (1)
Chromosomes: are normally visible under a light microscope only when the cell is undergoing
the metaphase of cell division. In eukaryotes, nuclear chromosomes are packaged by proteins
into a condensed structure called chromatin. Chromosomes must be replicated, divided, and
passed successfully to their daughter cells.

Unduplicated chromosomes are single double helixes, whereas duplicated chromosomes

contain two identical copies (called chromatids or sister chromatids) joined by a centromere.

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Definitions (2)

Homologous chromosomes: set of one

maternal and one paternal chromosome that
pair up with each other within a cell. These
copies have the same genes (but can have 2
different alleles) in the same loci.

Ploidy: number of sets of homologous

chromosomes in a cell.

Haploid cells (n) (sperms and eggs) have only

one set of chromosomes.

Diploid cells (2n) (somatic cells) have two sets

of chromosomes, derived from each parent.

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Ploidy

Diploid 2n: 2 sets of homologous chromosomes

Haploid n: 1 single set of homologous chromosomes

a1 b1 Diploid (2n): 2 sets of Haploid (n): 1 set of

homologous a1 homologous
chromosomes b1 chromosomes
a2 b2
4 chromatids 2 chromatids
4 chromosomes 2 chromosomes

a1 a1 b1 b1 Diploid (2n): 2 sets of Haploid (n): 1 copy of

duplicated homologous b1 each chromosome
b1
chromosomes
a2 a2 b2 b2 a1 a1 4 chromatids
8 chromatids 2 chromosomes
4 chromosomes

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Mitosis and meiosis

Mitosis: vital process during which the

fertilized egg (zygote) develops into a
mature organism: daughter cells are
diploid.

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Mitosis and meiosis

Mitosis: vital process during which the

fertilized egg (zygote) develops into a
mature organism: daughter cells are
diploid.

Meiosis: during the formation of

gametes, the number of chromosomes
is reduced by half: daughter cells are
haploid.

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Mitosis

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Mitosis

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 Mitosis
Mitosis is a part of the cell cycle in
which chromosomes in a cell nucleus are
separated into two identical sets of
chromosomes, and each set ends up in
its own nucleus

Mitosis is accompanied by cytokinesis,

which divides
the cytoplasm, organelles and cell
membrane into two new cells containing
roughly equal shares of these cellular
components

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Meiosis

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Meiosis
Meiosis is a specialized type of cell division that reduces the chromosome number by half
- Two phases: MEIOSIS I (reduction) and MEIOSIS II (division)

- Two kinds of reassortments in meiosis I give rise to genetic variation:

1. Crossing-over: exchanges of segments of homologous chromosomes; the process reassorts genes
on single chromosomes.
2. Random assortment of paternal and maternal homologs.

Crossing-over

Possible results of random assortment

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Analysis of cell proliferation

Use of radioactive thymidine:

- Identification of cells in S phase: cells

incorporate labeled thymidine exclusively
during DNA synthesis.
- Cells are analyzed by autoradiography
after the incorporation of thymidine.

1) Add thymidine to cells 2) Incubate cells 3) Eliminate the excess 4) Autoradiography

of thymidine from supernatant

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Analysis of the phases of cell cycle

Use of FCM to distinguish the phases of cell

cycle:

The advantages of FCM:

- Thousands of cells per sample can be
measured instead of only a few cells.
- Data analysis is rapid and
semiautomatic.
- FCM yields the data almost
instantaneously.

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Flow cytometry and cell cycle (1)

Cells at different stages of the cell cycle can

be distinguished by their DNA content
through FCM analysis.

1) Cells are permeabilized and labeled with a

fluorescent dye that binds the DNA:
propidium iodide (PI) intercalates between
bases, with no preference for purines or
pirimidines base pairs.

2) Cells are then passed through a FCM

device, which measures the fluorescence
intensity of single cells.

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Flow cytometry and cell cycle (2)

The data are plotted as cell number versus fluorescence intensity (A) (i.e. Single-parameter
histogram), which is proportional to DNA content (B)

A) B)

The amount of PI incorporated

is proportional to DNA content

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Thank you for your
kind attention!

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