SBS 2023

GENETICS

LECTURE 1
DNA : THE GENETIC MATERIAL

DR. MOHD NAZMI BIN ABD MANAP

 OBJECTIVES

 To trace the search for the genetic

material

 To describe the composition and

structure of DNA and RNA

 To describe the organization of DNA in

chromosomes
 Genetics: Subdisciplines
• Transmission genetics (classical genetics) –
traits are transmitted from generation to
generation

• Molecular genetics – molecular structure &

function of genes

• Population genetics – heredity in groups of

individuals for traits that are determined by one
or a few genes

• Quantitative genetics – heredity of traits in

groups of individuals that are determined by
many genes simultaneously
 Classical Genetics
Transmission of Genetic Information

Example of easily distinguishable alternative traits: purple-

flowered (left) vs. white-flowered (right) pea plants.
 Molecular Genetics
DNA, Chromosomes & Cell
 Molecular Genetics
DNA, Chromosomes & Cell

Stylized
diagram of the
relationship
between DNA,
chromosomes,
and the cell.
The Genetics Material: DNA
 DNA: The Genetic Material
Different DNA Structures

Different forms of DNA : (a) A-DNA. (b) B-DNA. (c) Z-DNA.

Genetic Map: Chromosome

Example of a genetic map, illustrating

some of the genes on chromosome 2 of the
fruit fly, Drosophila melanogaster. The
numerical values represent the positions of
the genes from the chromosome end (top)
measured in map units.
 Eukaryotic Cells

Eukaryotic cells. Cutaway diagrams of (a) a generalized higher plant cell and (b)
a generalized animal cell, showing the main organizational features and the
principal organelles in each cell
 Prokaryotic Cell

Cutaway diagram of a generalized prokaryotic cell.

 Gene Expression
Transcription and Translation
1st Gene Expression: Transcription
Expression of Genetic Information

Transcription. The DNA separates locally into single strands, and

RNA polymerase makes an RNA copy of one of the DNA
strands.
 2nd Gene Expression: Translation
Expression of Genetic Information

A ribosome, the organelle on which translation of mRNA (protein

synthesis) takes place. Two views of three-dimensional models
of the E. coli ribosome. The large subunit is red, and the small
subunit is yellow.
Lecture 1:

• Search for genetic material---nucleic acid or

protein/DNA or RNA?

 Griffith’s Transformation Experiment

 Avery’s Transformation Experiment
 Hershey-Chase Bacteriophage Experiment
 Tobacco Mosaic Virus (TMV) Experiment

• Nucleotides - composition and structure

• Double-helix model of DNA - Watson & Crick

• Organization of DNA/RNA in chromosomes

 Search for the Genetic Material
• Some substances must be responsible for passage of
traits from parents to offspring. Must have the
followings:

 Stability - stable enough to store information for long

periods

 Replication - able to replicate accurately

 Mutation - capable of change to allow evolution

 Variation - capable of almost infinite variation in structure

to account for all the different genes that exist

 Synthesis - able to cause the synthesis of all the kinds of

necessary molecules for development and metabolism
 Search for the Genetic Material
• Timeline of events:

• 1890 August Weismann proposed that a substance in the cell

nuclei controls the development of cells.

• 1900 Chromosomes shown to contain hereditary information,

later shown to be composed of protein & nucleic acids.

• 1928 Griffith’s Transformation Experiment

• 1944 Avery’s Transformation Experiment

• 1953 Hershey-Chase Bacteriophage Experiment

• 1953 Watson & Crick propose double-helix model of DNA

• 1956 Gierer & Schramm first demonstrate RNA as viral

genetic material.

• 1957 Fraenkel-Conrat & Singer confirm Gierer & Schramm

Frederick Griffith’s Transformation Experiment - 1928

“transforming principle” demonstrated with Streptococcus pneumoniae

Griffith hypothesized that the transforming agent was a “IIIS” protein.

 Griffith’s Transformation Experiment
Frederick Griffith’s 1928 experiment with Streptococcus pneumoniae
bacteria in mice showed that something passed from dead
bacteria (heredity from the dead) into nearby living ones,
allowing them to change their cell surface

Expt Bacteria type Effect on the mice Reasons

injected
1 Living type II R Harmless: mice R type bacteria do
survived not cause disease
2 Living type III S Virulent: mice died S type bacteria
cause disease

3 Heat-killed type III S Harmless: mice Dead bacteria do

survived not cause disease

4 Living type II R + Virulent: mice died Living III S

Heat-killed type III S recovered from
dead mice
He called this agent the Transforming Principle, but did not know what/how
Oswald T. Avery’s Transformation Experiment - 1944

 Lysed open IIIS bacteria cells and used the separated cell
components (cell extract)

 The extract was incubated with IIR bacteria cells. Colonies of

IIS bacteria grows from this culture plate.

 This shows that the ‘cell extract’ contains the transforming

principle

 Avery suggest that one of the component in this ‘cell extract’

could be the ‘transforming principle’

 The ‘cell extract’ contains:

 Polysaccharides
 Proteins
 DNA
 RNA
Oswald T. Avery’s Transformation Experiment - 1944

Determined that “IIIS” DNA was the genetic material responsible for
Griffith’s results (not RNA).
Hershey-Chase Bacteriophage Experiment - 1953

Bacteriophage = Virus that

attacks bacteria and
replicates by invading a
living cell and using the
cell’s molecular machinery.

Structure of T2 phage
Life cycle of virulent T2 phage:
Hershey-Chase Bacteriophage Experiment - 1953
 Hershey-Chase Bacteriophage Experiment - 1953

1. T2 bacteriophage is composed of DNA and proteins:

2. Set-up two replicates:

• Label DNA with 32P

• Label Protein with 35S

3. Infect E. coli bacteria with two types of labeled T2

4. Findings:
32P is discovered within the bacteria and progeny phages, whereas 35S is
not found within the bacteria but released with phage ghosts.

1969: Alfred Hershey

Gierer & Schramm Tobacco Mosaic Virus (TMV) Experiment - 1956
Fraenkel-Conrat & Singer - 1957

• Used 2 viral strains to demonstrate RNA is the genetic material of TMV

Nucleotide = monomers that make up DNA and RNA

Three components

1. Pentose (5-carbon) sugar

DNA = deoxyribose
RNA = ribose

2. Nitrogenous base

Purines
Adenine
Guanine

Pyrimidines
Cytosine
Uracil (RNA)
Thymine (DNA)

3. Phosphate group
Nucleotides are linked by phosphodiester bonds
to form polynucleotides.

phosphodiester bond :-
covalent bond between phosphate group of one
nucleotide and the 3’ carbon sugar of another
nucleotide (this bond is very strong and for this
reason DNA is remarkably stable).

5’ and 3’ end
One end of the chain has a 5’ carbon end and
the other end has a 3’ carbon
5’ end

3’ end
James D. Watson & Francis H. Crick - 1953

Double Helix Model of DNA

Two sources of information:

1. Base composition studies of Erwin Chargaff

• indicated double-stranded DNA consists of ~50% purines (A,G)

and ~50% pyrimidines (T, C)

• amount of A = amount of T and amount of G = amount of C

(Chargraff’s rules)

• %GC content varies from organism to organism

Examples: %A %T %G %C %GC

Homo sapiens 31.0 31.5 19.1 18.4 37.5

Zea mays 25.6 25.3 24.5 24.6 49.1
Drosophila 27.3 27.6 22.5 22.5 45.0
Aythya americana 25.8 25.8 24.2 24.2 48.4
James D. Watson & Francis H. Crick - 1953

Double Helix Model of DNA

Two sources of information:

2. X-ray diffraction studies of Rosalind Franklin & Maurice H. F.

Wilkins

Conclusion-DNA is a helical structure with

distinctive regularities, 0.34 nm & 3.4 nm.
Double Helix Model of DNA: Six main features

1. Two polynucleotide chains wound in a right-handed (clockwise)

double-helix.

2. Nucleotide chains are antiparallel: 5’  3’

3’  5’

3. Sugar-phosphate backbones are on the outside of the double

helix, and the bases are oriented towards the central axis.

4. Complementary base pairs from opposite strands are bound

together by relatively weak hydrogen bonds.

A pairs with T (2 H-bonds), and G pairs with C (3 H-bonds).

e.g., 5’-TATTCCGA-3’
3’-ATAAGGCT-3’

5. Base pairs are 0.34 nm apart. One complete turn of the helix
requires 3.4 nm (10 bases/turn).

6. Sugar-phosphate backbones are not equally-spaced, resulting

in major and minor grooves.
B-DNA
RNA (A pairs with U and C pairs with G)

Examples:
mRNA messenger RNA
tRNA transfer RNA
single-stranded
rRNA ribosomal RNA
snRNA small nuclear RNA

RNA secondary structure:

Yeast Alanine tRNA

Organization of DNA/RNA in chromosomes

Genome = chromosome or set of chromosomes that contains all the

DNA an organism (or organelle) possesses

Viral chromosomes 1. single or double-stranded DNA or RNA

2. circular or linear
3. surrounded by proteins
TMV T2 bacteriophage  bacteriophage

Prokaryotic chromosomes

1. most contain one double-stranded circular

DNA chromosome
2. others consist of one or more chromosomes
and are either circular or linear
3. typically arranged in arranged in a dense
clump in a region called the nucleoid
Problem:

Measured linearly, the Escherichia coli genome (4.6 Mb) would be

1,000 times longer than the E. coli cell.

The human genome (3.4 Gb) would be 2.3 m long if stretched

linearly.

Solutions:

1. Supercoiling DNA double helix is twisted in space about its

own axis, a process is controlled by
topoisomerases (enzymes).
(occurs in circular and linear DNA molecules)

2. Looped domains
More about genome size:

C value = total amount of DNA in the haploid (1N) genome

Varies widely from species to species and shows no relationship

to structural or organizational complexity.

Examples C value (bp)


48,502
T4
168,900
HIV-1
9,750
E. Coli
4,639,221
Lilium formosanum
36,000,000,000
Zea mays
5,000,000,000
Amoeba proteus
290,000,000,000
Eukaryotic chromosome structure

Chromatin complex of DNA and chomosomal proteins

~ twice as much protein as DNA

Two major types of proteins:

1. Histones abundant, basic proteins with a positive charge

that bind to DNA

5 main types: H1, H2A, H2B, H3, H4

~equal in mass to DNA

evolutionarily conserved

2. Non-histones all the other proteins associated with DNA

differ markedly in type and structure

amounts vary widely

>> 100% DNA mass
<< 50% DNA mass
Packing of DNA into chromosomes:

1. Level 1 Winding of DNA around histones to create a

nucleosome structure.

2. Level 2 Nucleosomes connected by

strands of linker DNA like
beads on a string.

3. Level 3 Packaging of nucleosomes into

30-nm chromatin fiber.

4. Level 4 Formation of looped domains.

Type of DNA Sequence:

•Centromeric DNA (CEN) Specialized sequences that function

for the accurate segregation of the
replicated chromosome to the
daughter cells during mitosis and meiosis

•Telomeric DNA At extreme ends of the chromosome,

maintain stability, and consist of
tandem repeats. Play a role in DNA
replication and stability of DNA.

•Unique-sequence DNA Often referred to as single-copy and

usually code for genes.

•Repetitive-sequence DNA May be interspersed or clustered and

vary in size.

 SINEs short interspersed repeated sequences (100-500 bp)

 LINEs long interspersed repeated sequences (>5,000 bp)

 Microsatellites short tandem repeats (e.g., TTA|TTA|TTA)

Text book price : RM100

Title: iGenetics, A
Molecular Approach