Eukaryotic Chromosomes

• Every species has a

different number of
chromosomes
• Humans have 46
chromosomes in 23
nearly identical pairs
– Additional/missing
chromosomes usually
fatal

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 Chromosomes
• Composed of chromatin – complex of DNA and
protein
• DNA of a single chromosome is one long
continuous double-stranded fiber
• Typical human chromosome 140 million
nucleotides long
• In the nondividing nucleus
– Heterochromatin – not expressed
– Euchromatin – expressed

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 Karyotype
• Particular array of chromosomes in an individual
organism
– Arranged according to size, staining properties,
location of centromere, etc.
• Humans are diploid (2n)
– 2 complete sets of chromosomes
– 46 total chromosomes
• Haploid (n) – 1 set of chromosomes
– 23 in humans
• Pair of chromosomes are homologous
– Each one is a homologue
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 Replication
• Prior to replication, each chromosome
composed of a single DNA molecule
• After replication, each chromosome
composed of 2 identical DNA molecules
– Held together by cohesin proteins
• Visible as 2 strands held together as
chromosome becomes more condensed
– One chromosome composed of 2 sister
chromatids

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 Eukaryotic Cell Cycle
1. G1 (gap phase 1)
– Primary growth phase, longest phase
2. S (synthesis) Interphase
– Replication of DNA
3. G2 (gap phase 2)
– Organelles replicate, microtubules organize
4. M (mitosis)
– Subdivided into 5 phases
5. C (cytokinesis)
– Separation of 2 new cells

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 Duration
• Time it takes to complete a cell cycle varies
greatly
• Fruit fly embryos = 8 minutes
• Mature cells take longer to grow
– Typical mammalian cell takes 24 hours
– Liver cell takes more than a year
• Growth occurs during G1, G2, and S phases
– M phase takes only about an hour
• Most variation in length of G1
– Resting phase G0 – cells spend more or less time here

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 Interphase
• G1, S, and G2 phases
– G1 – cells undergo major portion of growth
– S – replicate DNA
– G2 – chromosomes coil more tightly using motor
proteins; centrioles replicate; tubulin synthesis
• Centromere – point of constriction
– Kinetochore – attachment site for microtubules
– Each sister chromatid has a centromere
– Chromatids stay attached at centromere by cohesin

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 M phase
Mitosis is divided into 5 phases:
1. Prophase
2. Prometaphase
3. Metaphase
4. Anaphase
5. Telophase

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 Prophase
• Individual condensed chromosomes first become
visible with the light microscope
– Condensation continues throughout prophase
• Spindle apparatus assembles
– 2 centrioles move to opposite poles forming spindle
apparatus (no centrioles in plants)
– Asters – radial array of microtubules in animals (not
plants)
• Nuclear envelope breaks down

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 Prometaphase
• Transition occurs after disassembly of nuclear
envelope
• Microtubule attachment
– 2nd group grows from poles and attaches to
kinetochores
– Each sister chromatid connected to opposite poles
• Chromosomes begin to move to center of cell –
congression
– Assembly and disassembly of microtubules
– Motor proteins at kinetochores
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 Metaphase
• Alignment of
chromosomes
along metaphase
plate
– Not an actual
structure
– Future axis of cell
division

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 Anaphase
• Begins when centromeres split
• Key event is removal of cohesin proteins
from all chromosomes
• Sister chromatids pulled to opposite poles
• 2 forms of movements
– Anaphase A – kinetochores pulled toward
poles
– Anaphase B – poles move apart

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 Telophase
• Spindle apparatus disassembles
• Nuclear envelope forms around each set
of sister chromatids
– Now called chromosomes
• Chromosomes begin to uncoil
• Nucleolus reappears in each new nucleus

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 Cytokinesis
• Cleavage of the cell into equal halves
• Animal cells – constriction of actin
filaments produces a cleavage furrow
• Plant cells – cell plate forms between the
nuclei

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 Control of the Cell Cycle
• Current view integrates 2 concepts
1. Cell cycle has two irreversible points
– Replication of genetic material
– Separation of the sister chromatids
2. Cell cycle can be put on hold at specific points
called checkpoints
– Process is checked for accuracy and can be halted if
there are errors
– Allows cell to respond to internal and external signals

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 3 Checkpoints
1. G1/S checkpoint
– Cell “decides” to divide
– Primary point for external signal influence
2. G2/M checkpoint
– Cell makes a commitment to mitosis
– Assesses success of DNA replication
3. Late metaphase (spindle) checkpoint
– Cell ensures that all chromosomes are
attached to the spindle
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 Cyclin-dependent kinases (Cdks)
• Enzymes that phosphorylate proteins
• Primary mechanism of cell cycle control
• Cdks partner with different cyclins at different
points in the cell cycle
• For many years, a common view was that
cyclins drove the cell cycle – that is, the periodic
synthesis and destruction of cyclins acted as a
clock
• Now clear that Cdk itself is also controlled by
phosphorylation
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 Yeast

Anaphase
Promoting
Complex

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• Cdk – cyclin complex
– Also called mitosis-promoting factor (MPF)
• Activity of Cdk is also controlled by the pattern of
phosphorylation
– Phosphorylation at one site (red) inactivates Cdk
– Phosphorylation at another site (green) activates Cdk
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Control in multicellular eukaryotes
• Multiple Cdks control the cycle as opposed
to the single Cdk in yeasts
• Animal cells respond to a greater variety of
external signals than do yeasts, which
primarily respond to signals necessary for
mating
• More complex controls allow the
integration of more input into control of the
cycle
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Mammals

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 Growth factors
• Act by triggering intracellular signaling
systems
• Platelet-derived growth factor (PDGF) one
of the first growth factors to be identified
• PDGF receptor is a receptor tyrosine
kinase (RTK) that initiates a MAP kinase
cascade to stimulate cell division
• Growth factors can override cellular
controls that otherwise inhibit cell division
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 Cancer
• Unrestrained, uncontrolled growth of cells
• Failure of cell cycle control

• Two kinds of genes can disturb the cell

cycle when they are mutated
1. Tumor-suppressor genes
2. Proto-oncogenes

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 Tumor-suppressor genes
• p53 plays a key role in G1 checkpoint
• p53 protein monitors integrity of DNA
– If DNA damaged, cell division halted and repair
enzymes stimulated
– If DNA damage is irreparable, p53 directs cell to kill
itself
• Prevent the development of mutated cells
containing mutations
• p53 is absent or damaged in many cancerous
cells
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 Proto-oncogenes
• Normal cellular genes that become oncogenes
when mutated
– Oncogenes can cause cancer
• Some encode receptors for growth factors
– If receptor is mutated in “on”, cell no longer depends
on growth factors
• Some encode signal transduction proteins
• Only one copy of a proto-oncogene needs to
undergo this mutation for uncontrolled division to
take place
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 Tumor-suppressor genes
• p53 gene and many others
• Both copies of a tumor-suppressor gene
must lose function for the cancerous
phenotype to develop
• First tumor-suppressor identified was the
retinoblastoma susceptibility gene (Rb)
– Predisposes individuals for a rare form of
cancer that affects the retina of the eye

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• Inheriting a single mutant copy of Rb means the
individual has only one “good” copy left
– During the hundreds of thousands of divisions that
occur to produce the retina, any error that damages
the remaining good copy leads to a cancerous cell
– Single cancerous cell in the retina then leads to the
formation of a retinoblastoma tumor
• Rb protein integrates signals from growth factors
– Role is to bind important regulatory proteins and
prevent stimulation of cyclin or Cdk production

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 Sexual life cycle
• Made up of meiosis and fertilization
• Diploid cells
– Somatic cells of adults have 2 sets of
chromosomes
• Haploid cells
– Gametes have only 1 set of chromosomes
• Offspring inherit genetic material from 2
parents

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• Life cycles of sexually reproducing organisms
involve the alternation of haploid and diploid
stages
• Some life cycles include longer diploid phases,
some include longer haploid phases
• In most animals, diploid state dominates
– Zygote first undergoes mitosis to produce diploid cells
– Later in the life cycle, some of these diploid cells
undergo meiosis to produce haploid gametes

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 Features of Meiosis
• Meiosis includes two rounds of division
– Meiosis I and meiosis II
– Each has prophase, metaphase, anaphase, and
telophase stages
• Synapsis
– During early prophase I
– Homologous chromosomes become closely
associated
– Includes formation of synaptonemal complexes
• Formation also called tetrad or bivalents

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 Crossing over
• Genetic recombination between nonsister
chromatids
• Allows the homologues to exchange
chromosomal material
• Alleles of genes that were formerly on
separate homologues can now be found
on the same homologue
• Chiasmata – site of crossing over
– Contact maintained until anaphase I
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• During metaphase I, the paired homologues
move to the metaphase plate and become
oriented with homologues of each pair attached
to opposite poles of the spindle
– In mitosis, homologues behave independently
• During anaphase I, homologues are pulled to
opposite poles for each pair of chromosomes
– In mitosis, sister chromatids are pulled to opposite
poles

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• First meiotic division is termed the “reduction
division”
– Results in daughter cells that contain one homologue
from each chromosome pair
• No DNA replication between meiotic divisions
• Second meiotic division does not further reduce
the number of chromosomes
– Separates the sister chromatids for each homologue

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 The Process of Meiosis
• Meiotic cells have an • Meiosis I
interphase period that – Prophase I
is similar to mitosis – Metaphase I
with G1, S, and G2 – Anaphase I
phases – Telophase I
• After interphase, • Meiosis II
germ-line cells enter – Prophase II
meiosis I – Metaphase II
– Anaphase II
– Telophase II

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 Prophase I
• Chromosomes coil tighter and become visible,
nuclear envelope disappears, spindle forms
• Each chromosome composed of 2 sister
chromatids
• Synapsis
– Homologues become closely associated
– Crossing over occurs between nonsister chromatids
– Remain attached at chiasmata
• Chiasmata move to the end of the chromosome
arm before metaphase I
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 Metaphase I
• Terminal chiasmata hold homologues
together following crossing over
• Microtubules from opposite poles attach to
each homologue
– Not each sister chromatid
• Homologues are aligned at the metaphase
plate side-by-side
• Orientation of each pair of homologues on
the spindle is random
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 Anaphase I
• Microtubules of the spindle shorten
– Chiasmata break
• Homologues are separated from each other and
move to opposite poles
– Sister chromatids remain attached to each other at
their centromeres
• Each pole has a complete haploid set of
chromosomes consisting of one member of each
homologous pair
• Independent assortment of maternal and
paternal chromosomes 60
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 Telophase I
• Nuclear envelope re-forms around each
daughter nucleus
• Sister chromatids are no longer identical
because of crossing over (prophase I)
• Cytokinesis may or may not occur after
telophase I
• Meiosis II occurs after an interval of
variable length
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 Meiosis II
• Resembles a mitotic division
• Prophase II: nuclear envelopes dissolve
and new spindle apparatus forms
• Metaphase II: chromosomes align on
metaphase plate
• Anaphase II: sister chromatids are
separated from each other
• Telophase II: nuclear envelope re-forms
around 4 sets of daughter chromosomes;
cytokinesis follows 64
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 Final result
• Four cells containing haploid sets of
chromosomes
• In animals, develop directly into gametes
• In plants, fungi, and many protists, divide
mitotically
– Produce greater number of gametes
– Adults with varying numbers of gametes

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 Errors
• Nondisjunction – failure of chromosomes
to move to opposite poles during either
meiotic division
• Aneuploid gametes – gametes with
missing or extra chromosomes
• Most common cause of spontaneous
abortion in humans

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 Meiosis vs. Mitosis
• Meiosis is characterized by 4 features:
1. Synapsis and crossing over
2. Sister chromatids remain joined at their
centromeres throughout meiosis I
3. Kinetochores of sister chromatids attach to
the same pole in meiosis I
4. DNA replication is suppressed between
meiosis I and meiosis II
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