Principles of Genetics Analysis

(BIOL
3121)
Lauralee
Sherwood
Hillar Klandorf
Concord
Campus-Summer
2016
Paul Yancey

Lecture 13
Cell Division
08.11.16

Kip McGilliard Eastern Illinois University

Dr. Mazen Sidani

mazen.sidani@csueastbay.edu

Figure 11.11 Chromosome shape.

Karyotypes
A karyotype is a visual display of chromosomes as seen by
microscopy; it may be used to identify abnormalities in
chromosome number or structure
The chromosomes are grouped into homologous pairs in
descending order of size
The sex chromosomes are identified separately
Chromosomes may be stained with dyes to show distinctive
banding patterns for each chromosome
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In Situ Hybridization
In situ hybridization uses molecular probes, labeled with
fluorescence, to detect their target sequences
This approach is used to examine chromosomes with great
precision
First-generation methods used nucleotide probes labeled
with 32P

A human karyotype.

Fluorescent In Situ Hybridization

Currently, fluorescent in situ hybridization uses molecular
probes labeled with compounds that emit fluorescent light
when excited by UV or visible light
Various labels that emit light of different wavelengths can be
used simultaneously

For human chromosomes, 24 different

fluorophores are available that are unique to each
chromosome
There are also gene-specific fluorescent probes
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Fluorescent in situ hybridization (FISH).

Fluorescent in situ hybridization (FISH).

Imaging Chromosome Territory during

Interphase
Chromosomes are not uniformly distributed within a nucleus
Wilson, Sutton, and Boveri, who first observed this,
suggested that the variation in position might be related to
gene activity
Cremer and Cremer showed that chromosomes are
partitioned into specific regions, chromosome territories,
during interphase

Dynamic Chromosomes
Chromosomes do not occupy the same territory in each
nucleus, but once confined to a territory, a chromosome
does not leave until the M phase is initiated
However, chromosomes are active within their territories
and move, twist, and turn during transcription and DNA
replication
Chromosomes appear to be anchored in their territories by
their centromeres

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Dynamic Chromosomes (continued)

Interchromosomal domains are regions between territories

These are channels for movement of proteins,

enzymes, and RNA molecules
Early replicating parts of chromosomes are generally near
the center of the nucleus and late replicating parts are near
the periphery

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Chromosome territories in the eukaryotic nucleus.

Chromosome Position and Transcriptional

Activity
Transcriptionally active portions of chromosomes are found
nearer to interchromosomal domains, probably due to:
1. Greater access to needed proteins and enzymes
2. Faster dispersal of RNA transcripts, once they are
completed

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A three-dimensional model of chromosomes in the yeast

nucleus.

Chromosome Banding Techniques

Chromosome banding allows cytogeneticists to identify
each chromosome in a karyotype
Different stains and dyes are used to produce banding
patterns

The standard for human chromosome banding is

G (Giemsa) banding; the patterns are distinct
and reproducible

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Standardized human chromosome banding

patterns.

Heterochromatin and Euchromatin

Multiple genes can be contained in each chromosome
band
Chromosome condensation varies from one part of a
chromosome to another
Regions that contain actively expressed genes and are
less condensed during interphase are called
euchromatin
Regions that remain condensed in interphase and
contain many fewer expressed genes are called
heterochromatin
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Types of Heterochromatin
Facultative heterochromatin exhibits variable levels of
condensation, related to levels of transcription of resident
genes
Constitutive heterochromatin is permanently condensed,
found prominently in centromeres and telomeres, and
composed primarily of repetitive DNA sequences

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Replication and Transcription are Very Important

for Cell Division

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Cell Division
Mitosis produces two identical daughter cells that are exact
genetic replicas of the parental cell
Most body cells are somatic cells (non-reproductive),
usually with chromosomes present in pairs, which is the
diploid number (2n) of chromosomes
The haploid number of chromosomes includes one of each
chromosome pair (n)

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Reproductive Cells
Gametes are produced from germ-line, or reproductive, cells

Meiosis produces gametes that have half the number of

chromosomes as the original cell

The gametes are not genetically identical to one another

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Chromosomes
Sex chromosomes determine sex and differ between
genders
Certain processes equalize the expression of genes carried
on the sex chromosomes
Discovery of genes on the X chromosome supported the
chromosome theory of heredity

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3.1 Mitosis Divides Somatic Cells

Mitosis is the process of cell division that produces two
genetically identical daughter cells from one original parental
cell
It is precisely controlled to prevent either an excess or
insufficient number of cells

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Stages of the Cell Cycle

Cell division is regulated by control of the cell cycle, a cycle
of DNA replication and division
Cell cycles of all eukaryotes are similar
The two principal phases of the cell cycle are M phase, the
short time during which the cells divide, and a longer
interphase, the time between M phases

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The cell cycle.

Interphase
During the Gap 1 (G1) phase of interphase, all proteins
needed for normal cell function are transcribed and
translated; the duration of G1 varies
DNA is replicated during S phase or synthesis phase,
which follows G1
A small number of cells enter G0 after G1; cells in G0 never
progress through the cell cycle

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Interphase (continued)
During S phase, DNA replication results in doubling of the
DNA in each nucleus
Two sister chromatids are produced for each
chromosome
The completion of S phase leads into G2 or Gap 2 phase,
during which the cells prepare for division

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The cell cycle.

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Substages of M Phase
M phase is divided into
Prophase
Prometaphase
Metaphase
Anaphase
Telophase
M phase accomplishes karyokinesis, partitioning of DNA
into daughter cell nuclei, and cytokinesis, the partitioning of
the cytoplasm

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Chromosomes during Mitosis

Cells at the beginning and the end of mitosis are diploid (2n)
Progressive condensation of chromosomes begins in
prophase and reaches a maximum in metaphase
Centromeres, specialized sequences where sister
chromatids are joined together, become visible in prophase;
centromeres bind protein complexes called kinetochores

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Interphase and the five stages of mitosis.

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Interphase and the five stages of mitosis.

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Chromosome Distribution
In animal cells, two centrosomes appear, which migrate to
form the opposite poles of the dividing cell
Centrosomes are the source of spindle fiber microtubules;
microtubules have a minus () end at the centrosome and a
plus () end that grows away from the centrosome
The spindle fibers emanate from the centrosomes in a
pattern called the aster

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Microtubules in dividing cells emanate from centrosomes.

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Types of Microtubules in Cells

Kinetochore microtubules embed in the
kinetochore at the centromere of each chromatid,
and are responsible for chromosome movement
Polar microtubules extend toward the opposite
pole of the centrosome and contribute to cell
elongation and cell stability
Astral microtubules grow toward the membrane
of the cell and contribute to cell stability

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Metaphase Chromosomes
By the end of prometaphase, kinetochore microtubules are
bound to each kinetochore
Metaphase chromosomes are condensed10,000-fold
compared to the onset of prophase; these chromosomes are
pulled toward each centrosome by the kinetochore
microtubules
The opposing forces align the chromosomes along the
metaphase plate

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Sister Chromatid Cohesion

The tension created by the pull of the kinetochore
microtubules is balanced by sister chromatid cohesion
The protein cohesin localizes between sister chromatids and
holds them together, preventing their premature separation

Cohesin is a 4-subunit protein that coats sister chromatids

along their entire length, with the greatest concentration at
the centromeres

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Sister chromatid cohesion during mitosis.

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Anaphase
Sister chromatids separate at anaphase
and begin to move toward opposite poles
in the cell
In anaphase A the sister chromatids separate due to the
enzyme separase cleaving the polypeptides in cohesin
The separation of sister chromatids is called chromosome
disjunction

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Anaphase (continued)
During anaphase B, polar microtubules extend in length

This causes the cell to take on an elongated shape

The altered shape facilitates cytokinesis at the end of

telophase, leading to formation of two daughter cells

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Completion of Cell Division

In telophase, nuclear membranes reassemble around the
chromosomes at each pole

Decondensation returns chromosomes to their

diffuse interphase state
Two identical nuclei occupy the elongated cell, which will
divide into two daughter cells by cytokinesis

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Cytokinesis
In animal cells, a contractile ring of actin creates a cleavage
furrow around the circumference of the cell; this pinches the
cell in two
In plants, a new cell wall is constructed along the cellular
midline
In both plant and animal cells, cytokinesis divides the
cytoplasm and organelles between the daughter cells

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Cytokinesis in animal cells (a) and plant cells (b).

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Mitosis Produces Identical Daughter Cells

Mitosis separates replicated copies of sister chromatids into
identical nuclei, forming two genetically identical daughter
cells

The diploid number of chromosomes (2n) is maintained

throughout the cell cycle

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An overview of
mitosis.

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Cell Cycle Checkpoints

Common, genetically controlled signals drive the cell cycle
Cell cycle checkpoints are monitored by protein interactions
for readiness to progress to the next stage
A common mechanism is carried out by protein complexes
joining a protein kinase with a cyclin protein

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Cell cycle
checkpoints
and cyclin
proteins.

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Meiosis Produces Gametes for Sexual

Reproduction
Reproduction can be divided into two broad categories

In asexual reproduction, organisms reproduce without mating

and produce genetically identical offspring
In sexual reproduction, gametes (reproductive cells) are
produced; these unite during fertilization

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Multicellular Eukaryotes Reproduce Mainly

Sexually
Males and females carry distinct reproductive tissues and
structures
Mating requires the production of haploid gametes from both
male and female
The union of haploid gametes produces diploid progeny

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Meiosis versus Mitosis

Many features of meiosis are similar or identical to mitosis,
for example, interphase
Meiosis is distinguished from mitosis on the basis of events
during meiotic M phase and the production of four haploid
gametes
Meiotic interphase is followed by two division stages called
meiosis I and meiosis II with no DNA replication between
them

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Comparison of Mitosis and Meiosis

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Meiosis I and II
In meiosis I homologous chromosomes separate
from one another, reducing the diploid number
of chromosomes to the haploid number
In meiosis II, sister chromatids separate from
one another to produce four haploid gametes,
each with one chromosome of the original
diploid pair

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An overview of
meiosis.

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Meiosis I
Three hallmark events occur in meiosis I:
1. Homologous chromosome pairing
2. Crossing over between homologous chromosomes
3. Segregation (separation) of homologous chromosomes,
which reduces chromosomes to the haploid number

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Stages of Meiosis I
Meiosis I is divided into prophase I, metaphase I, anaphase I,
and telophase I
Pairing and recombination of homologs takes place in
prophase I
Prophase I is subdivided into five stages:
leptotene, zygotene, pachytene, diplotene, and diakinesis
(Laras Zebra is Playing Drums and Dancing)

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Leptotene and Zygotene

Chromosome condensation begins in leptotene stage
The meiotic spindle forms as microtubules extend out from
centrosomes
The nuclear envelope disintegrates during zygotene
Homologous chromosomes undergo synapsis

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Synaptonemal Complex
Homologous chromosomes align and the synaptonemal
complex (protein bridge) is formed between them
This tri-layer protein structure tightly binds nonsister
chromatids of homologous chromosomes
Nonsister chromatids belong to different members of a
homologous pair

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The synaptonemal complex.

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Figure 3.10 The stages of meiosis.

2015 Pearson

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Pachytene
Chromosome condensation continues in pachytene
Paired homologs are called tetrads, due to the four visible
chromatids
Recombination nodules can be seen at intervals in the
synaptonemal complex
These are aggregates of enzymes and proteins needed for
crossing over between homologs

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Diplotene
Chromosomes continue to condense in diplotene, and the
synaptonemal complex begins to dissolve
Homologs pull apart slightly, revealing chiasma at locations
where crossing over has occurred
Cohesin protein is present between sister chromatids, to
resist the pulling forces of kinetochore microtubules

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Diakinesis
Kinetochore microtubules move synapsed chromosome pairs
toward the metaphase plate

Here, homologs align side by side

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Homolog separation in meiosis I.

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Figure 3.10 The stages of meiosis.

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Metaphase I
In metaphase I chiasmata between homologs are dissolved;
this completes crossing over
Homologs align on opposite sides of the metaphase plate
Kinetochore microtubules attach to both sister chromatids of
one homolog; kinetochore microtubules from the opposite
pole do the same for the other homolog

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Anaphase I
Anaphase I begins when homologs separate from one
another and are pulled to opposite poles of the cell
Sister chromatids are firmly attached by cohesin

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Telophase I and Cytokinesis

In telophase I the nuclear membranes re-form around the
separated haploid sets of chromosomes

Cytokinesis follows telophase I and divides the cytoplasm to

create two haploid cells

Meiosis I is called the reduction division

because the ploidy of the daughter cells is
halved compared to the original diploid parent
cell
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Figure 3.10 The stages of meiosis.

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Meiosis II
Meiosis II divides each haploid daughter cell into two haploid
cells, by separating sister chromatids from one another

The process is similar to mitosis in a haploid cell

Four genetically distinct haploid cells are produced, each

carrying one chromosome of a homologous pair

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Figure 3.10 The stages of meiosis.

2015 Pearson

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 2015 Pearson

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 2015 Pearson

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 2015 Pearson

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The Mechanistic Basis of Mendelian Ratios

Separation of homologs and sister chromatids in meiosis
constitutes the mechanical basis of Mendels laws
For example, in an organism that is genotype Aa, the
homologs bearing A and a separate from one another during
anaphase I
At the end of meiosis, two gametes have the A allele and two
have a; this generates the 1:1 ratio predicted by the law of
segregation

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The Mechanistic Basis of Mendelian Ratios

Separation of homologs and sister chromatids in meiosis
constitutes the mechanical basis of Mendels laws
For example, in an organism that is genotype Aa, the
homologs bearing A and a separate from one another during
anaphase I
At the end of meiosis, two gametes have the A allele and two
have a; this generates the 1:1 ratio predicted by the law of
segregation

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Meiosis
and the
law of segregation.

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Independent Assortment
Independent assortment of alleles is illustrated by behavior of
two pairs of homologs during meiosis
For an organism with genotype AaBb, two equally likely
arrangements of paired homologs can occur
One yields gametes AB and ab, whereas the other produces
gametes Ab and aB; these four gamete combinations occur
with equal likelihood

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Meiosis and the law of independent assortment.

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The Chromosome Theory of Heredity Proposes That

Genes Are Carried on Chromosomes
Sutton and Boveri proposed that chromosome behavior in
meiosis mirrors hereditary transmission of genes
Morgan studied fruit flies, Drosophila melanogaster, to test
Mendels rules on a natural, rather than domesticated,
species
The term wild type signifies the phenotype most common in
a population

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Morgans Results
Over several years, Morgans lab members found
numerous phenotypic variants and analyzed these in
controlled crosses

He concluded from his results that genes

were carried on chromosomes

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X-Linked Inheritance
After studying insects, Nettie Stevens concluded that sex-

dependent hereditary differences are due to the

presence of two X chromosomes in females and
an X and a smaller Y chromosome in males
Sex-linked inheritance refers to transmission of genes on
sex chromosomes
In 1910, Morgans experiments with flies validated Stevenss
conclusions

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X and Y Chromosomes
Stevens studied chromosome differences between male and
female beetles (Tenebrio molitor)
She found that diploid cells of females contained 20 large
chromosomes but those of males contained 19 large
chromosomes and one small one
Half of the sperm of males contained 10 large chromosomes
and the other half had 9 large and one small chromosome

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3.4 Sex Determination Is Chromosomal and

Genetic
Sex determination involves the genetic and biological
processes that produce the male and female characteristics
of a species
Chromosomal sex is the presence of chromosomes
characteristic of each sex and is determined at the moment
of fertilization
Phenotypic sex is the internal and external morphology of
each sex, and results from differences in gene expression

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The X/Autosome Ratio

The X/A ratio or X/autosome ratio determines gender
based on the number of X chromosomes to sets of
autosomes

Males have an X/A ratio of 0.5 and

females have a ratio of 1.0

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Mammalian Sex Determination

Placental mammals have X and Y chromosomes too
Sex determination depends on the presence or absence
of a single gene, SRY, found on the Y chromosome

In mammals, males can be XY (normal),

XXY, or XYY; females are XX (normal), XO,
or XXX
SRY produces a transcription factor needed for male-specific
gene expression
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SRY
Early mammalian embryos have clusters of tissue called
undifferentiated gonads, which can develop as ovaries or
testes
Expression of SRY initiates testicular development of the
undifferentiated gonads
The absence of SRY expression allows the default state,
female, to develop

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Mammalian sex determination is initiated by the Y-linked

SRY gene.

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Diversity of Sex Determination

A different system, the Z/W system, is used by birds,

some reptiles, some fish, butterflies, and moths

In this system females have two different sex
chromosomes (ZW), and males have two sex
chromosomes that are the same (ZZ)
Sex chromosomes of the platypus consist of five pairs of sex
chromosomes with five XY pairs in males and five XX pairs in
females

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ZW inheritance of feather form in poultry is revealed by

analysis of reciprocal crosses.

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Figure 3.22b ZW inheritance of feather form in poultry is

revealed by analysis of reciprocal crosses.

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