LES106 Biology: Form and Function

Semester 1, 2023/2024
Week 3

Sammy Chan
Module Instructor
 This Lecture
• Bacterial cell division (binary fission)
• Eukaryotic chromatin packaging
• Ploidy and Karyotype
• Eukaryotic cell cycle
• Eukaryotic cell division
• Mitosis – growth, repair, regeneration
• Meiosis (next lecture)
 Learning Objectives

• Describe process of binary fission

– What is final outcome of this process?
 Cell Division
• Prokaryotic and eukaryotic cells
– Produces 2 new cells with the same genetic information as the original

• Duplication and segregation of genetic material

• Bacteria divide as a way of reproducing themselves

 Bacterial Cell Division
• Bacteria divide by binary fission
– No sexual cycle
– Asexual reproduction: used by single-cell organisms, usually
prokaryotes
– Reproduction is clonal (all cells identical to parent)
• Bacterial genome
– single, circular chromosome tightly packed in the cell at the
nucleoid region
• New chromosomes are partitioned to opposite ends of the cell
– Occurs when the cell elongates (grows)
• Septum forms to divide the cell into 2 cells
– A process called septation
– Via protein FtsZ
 Binary Fission 1. Prior to cell division, the bacterial DNA
1. molecule replicates. The replication of the
double-stranded, circular DNA molecule
that constitutes the genome of a
bacterium begins at a specific site, called
the origin of replication (green area).
a) Bacterial chromosomes
replicates from the
origin of replication 2.
2. The replication enzymes move out in
both directions from that site and make
copies of each strand in the DNA
duplex. The enzymes continue until
b) Enzymes replicates the they meet at another specific site, the
terminus of replication (red area).
chromosome

c) Cell elongates, DNA 3. 3. As the DNA is replicated, the cell

elongates, and the DNA is partitioned
partitioned to ends in the cell such that the origins are at
the ¼ and ¾ positions in the cell and
the termini are oriented toward the
middle of the cell.

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Binary Fission

4. FtsZ 4. Septation then begins, in which

new membrane and cell wall material
d) Septum begin to grow and form a septum at
approximately the midpoint of the
production via FtsZ cell. A protein molecule called FtsZ
facilitates this process.
Septum

5.
5. When the septum is complete, the
e) End result: cell pinches in two, and two daughter
2 identical cells cells are formed, each containing a
bacterial DNA molecule.

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
 Eukaryotic chromosomes
Learning Objectives

• Describe the structure of eukaryotic chromosomes.

• Distinguish between homologues and sister chromatids.

• Contrast replicated and nonreplicated chromosomes.

 Eukaryotic Chromosomes
• Every species has a different
number of chromosomes Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

• Chromosomes are usually linear

• Humans have 46 chromosomes

(23 pairs)
– Additional/missing chromosomes
usually fatal with some exceptions © Biophoto Associates/Photo Researchers, Inc. 950x
 Chromatin
• Chromosomes are composed of chromatin
– complex of DNA and protein
• Typical human chromosome 140 million (1.4 × 108)
nucleotides long

• A typical human cell is 20-30 µm

• 1 DNA molecule is 5 cm
• Full set of DNA >2 meters inside a diploid human nucleus
→ it must be compacted to fit inside the nucleus!
 Chromatin packaging

• From DNA to Chromosomes

1) Naked DNA
2) Nucleosome
3) Solenoid
4) Chromatin Loops
5) Chromosome
 Chromatin
• Nucleosome
– The complex of DNA and histone proteins is termed a nucleosome
– DNA duplex tightly coiled around histone proteins
– 8 histone proteins every 200 nucleotides
– Histones are positively charged and strongly attracted to negatively charged
phosphate groups of DNA Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

DNA Double Helix (duplex) Nucleosome

Histone core
DNA

+++

---
 Chromatin
• Nucleosomes wrapped into higher order coils called solenoids
– Leads to a fiber 30 nm in diameter
– Found in non-dividing (interphase) chromatin
• During mitosis, chromatin in solenoid arranged around scaffold of protein to
achieve maximum compaction
Chromosome Interphase chromatin
seen in dividing cells Chromatin in non-dividing cells

https://cytochemistry.net/cell-biology/nucleus2.htm
Eukaryotic Chromatin Organization

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
 Chromatin
• 2 types of chromatin
o Heterochromatin – condensed, genes not expressed
o Euchromatin – open, genes can be expressed
Heterochromatin
closed chromatin,
not expressed

Euchromatin
open chromatin,
expressed

https://medcell.org/
histology/cell_lab/eu
chromatin_and_hete
rochromatin.php
 Ploidy
• Defining the no. of different chromosomes in a species:
Haploid (n): 1 complete set of chromosomes necessary to define an organism
• Humans are diploid (2n)
– Each cell contains 2 complete sets of chromosomes
– Total no. of chromosomes = 2n = 46
– i.e. diploid number = 46, haploid number = 23
 Polyploidy
• Heritable condition of having more than 2 complete sets of chromosomes
– Common in plants

Bharadwaj, D.N. (2015). Polyploidy in Crop Improvement and Evolution. In: Bahadur, B., Venkat Rajam,
M., Sahijram, L., Krishnamurthy, K. (eds) Plant Biology and Biotechnology. Springer, New Delhi.
Chromosome number in different organisms
 Karyotype
• Particular array of chromosomes in an individual organism is called karyotype
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

• How many chromosomes are there?

• Is this cell haploid or diploid? Why?

© CNRI/Photo Researchers, Inc. 500x

 Karyotype

Turner’s syndrome Down syndrome

• Missing one chromosome: monosomy • Having an extra chromosome: trisomy
Embryos do not survive in most cases Usually fatal, with some exceptions

https://www.differencebetween.com/difference-between-monosomy-and-vs-trisomy/
 Homologous Chromosomes
• Chromosome pairs are homologous
– Maternal chromosome and paternal chromosome
– Each one is a homologue

What is an example of a pair of

homologous chromosomes?
 Learning Objectives

• Cell Cycle
– Describe the eukaryotic cell cycle

• Interphase
– Describe the events that take place during interphase
– Illustrate the connection between sister chromatids after S phase
 Eukaryotic Cell Division
The eukaryotic cell cycle has two phases:
Mitosis and cytokinesis
• eukaryotic nuclear division (mitosis) and cell division (cytokinesis)
Interphase
• Cell is being a cell
• Cell nucleus is visible
• Cell metabolic functions, including DNA replication, occur
• Begins after cytokinesis and ends when mitosis starts
 Eukaryotic Cell Cycle
1. G1 (gap phase 1)
Primary growth phase, longest phase
2. S (synthesis)
Replication of DNA Interphase
3. G2 (gap phase 2)
Organelles replicate, microtubules organize
4. M (mitosis)
– Subdivided into 5 phases
5. C (cytokinesis)
– Separation into 2 new cells
 The Cell Cycle M Phase

Metaphase
Prometaphase Anaphase
Prophase Telophase

G2: chromosomes G2
coil more tightly

G1
S Interphase
G2 G1: cells undergo
Mitosis
M Phase
major portion of
S: DNA replicates = Cytokinesis growth
two sister S G1
chromatids attached
at centromere
 Duration of Cell Cycle
• Most variation in the length of the cell cycle
between organisms or cell types occurs in G1
– Cells often pause in G1 before DNA replication, enters
a resting state called G0 M

– Resting phase G0 – cells spend more or less time here

before resuming cell division. G2

– Most of cells in animal’s body are in G0 phase

– Muscle and nerve cells remain there permanently
– Liver cells can resume G1 phase in response to factors S G1
released during injury

G0: resting phase

 Interphase: Preparation for Mitosis

G1, S, and G2 phases

• G1: cells undergo major portion of growth
• S: replicate DNA produce two sister chromatids attached at the
centromere
• G2: chromosomes coil more tightly using motor proteins; centrioles
replicate; tubulin synthesis
 The Cell Cycle M Phase

Metaphase
2nd growth phase. Prepare for
Prometaphase Anaphase
separation of newly replicated
DNA. Microtubules begin to Prophase Telophase
reorganise to form a spindle.
G2: chromosomes G2
coil more tightly

G1
S Interphase
G2 G1: cells undergo
Mitosis
M Phase
major portion of
S: DNA replicates = Cytokinesis growth
two sister S G1
chromatids attached Most cells: longest
at centromere phase
Synthesise a replica of
the genome
 Interphase: S phase
S: replicate DNA

two sister chromatids

attached at the centromere

Note:
Chromosomes still
not visible during
interphase
 Chromosome Replication
• Chromosome seen in karyotype only present for a brief period during cell division

Homologous Chromosomes Homologous Chromosomes After replication:

each chromosome
composed of 2
Before replication:
identical DNA
each chromosome
molecules
composed of a Kinetochore
- Sister chromatids
single DNA
held together at
molecule
Replication the centromeres
by cohesin proteins
Cohesin
Centromere
proteins
Kinetochores

*Only visible as 2 strands

held together when
chromosomes become
Sister chromatids condensed at the start of
Sister chromatids mitosis
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
 Kinetochores

• Centromere – point of
Sister
Cohesin chromatids constriction
proteins

Centromere
• Kinetochore proteins
region of
chromosome
assemble. Attachment site for
microtubules
• Each sister chromatid has
a centromere
Kinetochore
Kinetochore
• Chromatids stay attached at
microtubules centromere by cohesin
Metaphase
chromosome

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
 Centrioles
Interphase G2 (replicated;
animal
Chromatin
(replicated)
cells only)

G2: Aster
• Second growth phase
• Prepare to separate newly replicated
chromosomes
• Chromosomes coil more tightly (not
visible as (“X” shape yet)
Nuclear Nucleolus
membrane Nucleus

• DNA has been replicated

• Centrioles replicate (animal cells)
• Cell prepares for division

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
 Learning Objectives
• Describe the phases of mitosis.
– PPMAT

• Explain the importance of metaphase

• Compare cytokinesis in plants and animals

 Mitosis
Chromosome Segregation and the Division of Cytoplasmic Contents

Mitosis is divided into 5 phases: PPMAT

1. Prophase
2. Prometaphase
3. Metaphase
4. Anaphase
5. Telophase
Mitosis is divided into 5 phases: PPMAT
 Mitosis: Prophase
• Chromosomes condense and become visible

• Chromosomes appear as two sister chromatids held

together at the centromere Mitotic spindle Condensed
beginning to form chromosomes

• Nuclear envelope breaks down

• Mitotic spindle begins to form

– 2 centrioles move to opposite poles forming
spindle apparatus (no centrioles in plants)
– Asters – radial array of microtubules in animals
(not plants)
Prophase
• Golgi and ER are dispersed
 Mitosis: Prometaphase

• After disassembly of nuclear envelope

Centromere and Mitotic
kinetochore spindle
• Chromosomes attach to microtubules at the
kinetochores from opposite poles

• Chromosomes move to equator of the cell

• Motor proteins at kinetochores

Prometaphase
 Mitosis: Metaphase
Chromosomes
• Alignment of aligned on Kinetochore
chromosomes along metaphase plate microtubule
metaphase plate
– Imaginary structure
57µm
Polar
– Axis of cell division microtubule
Centrioles
Kinetochore
microtubule
• Chromosomes are
attached to opposite
poles and are under Polar microtubule
tension Aster
Metaphase
Metaphase plate Sister chromatids

© Andrew S. Bajer, University of Oregon

 Mitosis: Anaphase

• Begins when centromeres split

Polar
microtubule
Chromosomes
• Removal of cohesin proteins from all
chromosomes

• Sister chromatids pulled to opposite

poles

Kinetochore
microtubule

Anaphase
 Mitosis: Telophase

• Chromosomes are clustered at opposite poles

and decondense Kinetochore
Nucleus reforming microtubule
• Spindle apparatus disassembles

• Nuclear envelopes re-forms around

chromosomes

• Chromosomes begin to uncoil

Polar microtubule
• Nucleolus reappears in each new nucleus

• Golgi complex and ER re-form Telophase

 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
 Cytokinesis in Animal Cell: Animal cell cleavage

Contraction of actin filaments by

myosin pull the “equator” of the
cell causing an invagination of
the cell (a fissure called a
cleavage furrow).

The cleavage furrow gets

continuously deeper until it
eventually divides the cell into
cleavage two new, independent daughter
furrow cells.

a. 325 µm b. 25 µm
a: © David M. Phillips/Visuals Unlimited; b: © Guenter Albrecht-Buehler, Northwestern University, Chicago
 Cytokinesis in Plant Cell

A mixture of enzymes, proteins, and glucose

molecules are transported via vesicles to the
center of the cell.
19,000×
These vesicles continuously build upon each
Vesicles containing Cell wall other until a completely new cell wall has
membrane components
fusing to form cell plate emerged and two new cells are formed,
independent of one another.

(top): © E.H. Newcomb & W.P. Wergin/Biological Photo Service

Mitosis and Cytokinesis: Summary
 Outcome of Mitosis
• After Mitosis and Cytokinesis
– Two identical daughter cells
– Number and types of chromosomes remains the same
• 2n→ mitosis → 2n
• n → mitosis → n
• Used for
– Asexual reproduction
– Growth
– Repair
Example: Cell is 2n and we are looking at
chromosome 1 only (2 copies)

Interphase S-Phase
 Mitosis

Prophase - Metaphase Anaphase Telophase

Prometaphase
Cytokinesis
Try drawing the steps your self

Label:
Sister Chromatids, Spindles,
Centrioles, Chromosomes,
Diploid cells
 DNA Division in Different Organisms
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Prokaryotes Some Protists Other Protists Yeasts Animals

No nucleus, usually
have single circular
chromosome. After DNA
is replicated, it is
partitioned in the cell.
After cell elongation,
FtsZ protein assembles
into a ring and facilitates
septation and cell
division.
Chromosome
FtsZ protein
Nucleus present and
nuclear envelope
remains intact during
cell division.
Chromosomes line up.
Microtubule fibers pass
through tunnels in the
nuclear membrane and
set up an axis for
separation of replicated
chromosomes, and cell
division.
A spindle of micro-
tubules forms between
two pairs of centrioles at
opposite ends of the
cell. The spindle passes
through one tunnel in
the intact nuclear
envelope. Kinetochore
microtubules form
between kinetochores
on the chromosomes
and the spindle poles
and pull the chromo-
somes to each pole.
Nuclear envelope
remains intact; spindle
microtubules form
inside the nucleus
between spindle pole
bodies. A single
kinetochore microtubule
attaches to each
chromosome and
pulls each to a pole.
Kinetochore microtubule
Spindle pole body
Spindle microtubules
begin to form between
centrioles outside of
nucleus. Centrioles move
to the poles and the
nuclear envelope breaks
down. Kinetochore
microtubules attach
kinetochores of
chromosomes to spindle
poles. Polar microtubules
extend toward the center
of the cell and overlap.

Microtubule Kinetochore microtubule

Kinetochore microtubule
Chromosome Fragments
Central spindle of nuclear
of microtubules envelope

Septum

Polar microtubule

Nucleus

Centrioles Kinetochore Centriole Polar microtubule

 Control of the Cell Cycle
Current view integrates 2 concepts
1. Cell cycle has two irreversible points
I. Replication of genetic material
II. 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
 Control of the Cell Cycle
3 Checkpoints:
1. G1/S checkpoint G2/M checkpoint Spindle checkpoint

– Cell “decides” whether or not to divide

M
2. G2/M checkpoint
G2
– Cell makes a commitment to mitosis
– Assesses success of DNA replication
– Can stall the cycle if DNA has not been
accurately replicated G1/S checkpoint
G1
S (Start or restriction point)
3. Late metaphase (spindle) checkpoint
– Cell ensures that all chromosomes are
attached to the spindle
 3 Checkpoints
G2/M checkpoint Spindle checkpoint

G2

G1/S checkpoint
(Start or restriction point) G1
S
 Control of the Cell Cycle

Cyclin-dependent kinases (Cdks):

• Enzyme kinases that phosphorylate proteins
• Primary mechanism of cell cycle control
• Cdks partner with different cyclins at different points in the cell cycle
• Cdk itself is also controlled by phosphorylation
 Cyclin-dependent kinase P
(Cdk)

MPF
Cyclin

• 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
Checkpoints of the Mammalian Cell Cycle
G2/M Checkpoint

Cdk1/Cyclin B

• Replication
completed
• Cdk partners with different
• DNA integrity Cyclins (E or B) to control the
G1/S or G2 M Checkpoints
M

G2 • Cdk2 active or inactive based on

G1/S Checkpoint phosphorylation patterns
Cdk2/Cyclin E

• Growth factors
• Nutritional state
of cell G1
S
• Size of cell
 Checkpoints of the Mammalian Cell Cycle
Spindle Checkpoint
• APC (anaphase promoting complex)
APC
marks securin for destruction
• Chromosomes
attached at

• No securin → cohesion destroyed → metaphase plate

sister chromatids separate

M

G2

G1
S
 Cancer
Cancer:
Unrestrained, uncontrolled growth of cells
• Failure of cell cycle control
 Question 1
Prokaryotic cells divide by —
a. Mitosis
b. Cytokinesis
c. Binary fission
d. Replication
e. Conversion
 Question 2
The unexpressed form of DNA is _____; the expressed form of DNA is ______.
a. Heterochromatin; homochromatin
b. Euchromatin; mesochromatin
c. Mesochromatin; homochromatin
d. heterochromatin; euchromatin
e. None of the above
 Question 3
Homologous chromosomes and sister chromatids are the same thing.
a. This is true
b. This is false
 Question 4
Where would a researcher find histones?
a. In chromosomes with DNA coiled around them
b. In the spindle apparatus
c. At the formation of the cell plate
d. Surrounding a nuclear pore
e. Bound to a ribosome
 Question 5
What happens during the S phase of the cell cycle?
a. Growth and maturation
b. Replication of nuclear DNA
c. Production of extra organelles
d. Separation of chromosomes into sister chromatids
 Question 6

Mature neurons can spend the entire animal’s lifetime in

—
a. G0
b. G1
c. G2
d. S
e. M
 Question 7
If a researcher looked at a cell and noticed a straight line of sister chromatids, which
phase would they be looking at?
a. Prophase
b. Metaphase
c. Anaphase
d. Telophase
e. Interphase
 Question 8
Cytokinesis is the division of the nucleus, and mitosis is the division of the
cytoplasm.
a. This is true
b. This is false