1

CENTRAL DOGMA
Prepared by:
DYAN B. JUMAMOY
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CENTRAL DOGMA
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Discovering Nucleic Acids

Friedrich Miescher
 1869: isolated the genetic material
from WBC nuclei; he noted it had
an acidic nature and called it
nuclein

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Discovering the DNA Structure

Phoebus Levene
 1909: 5-carbon sugar ribose
 1929: discovered deoxyribose
 Discovered that the 3 parts of
nucleic acid are present in equal
proportions (phosphate-sugar-base
units called nucleotides)

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Discovering the DNA Structure

Frederick Griffith
 1928: studied the epidemiology
and pathology of 2 strains of
Streptococcus pneumoniae & took
the first step in identifying DNA as
the genetic material

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Discovering the DNA Structure

Oswald Avery, Colin MacLeod,

& Maclyn McCarty
 1944: explained further the result of
Griffith’s experiment
 They suggested that DNA, rather
than protein, may be the
hereditary material of bacteria –
and perhaps in higher organisms as
well
vle.du.ac.in
 7
Discovering the DNA Structure
Erwin Chargaff
 1950s
 DNA in several species
contains equal
amounts of the bases
A and T and equal
amounts of the bases
G and C
 Chargaff’s rule:
Adenine = Thymine
and Cytosine =
Guanine
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Discovering the DNA Structure

Alfred Hershey & Martha Chase

1952: used Escherichia coli
infected with bacteriophage
DNA is the genetic material and
not protein

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Discovering the DNA Structure

Maurice Wilkins & Rosalind

Franklin
Bombarded DNA with X-rays (X-
ray diffraction), then deduced
the overall structure of the
molecule from the patterns in
which the X-rays were deflected
10 Discovering the DNA Structure

Rosalind Franklin
 Distinguished 2 forms of DNA:
“A form” – dry, crystalline
“B form” – wetter type
Linus Pauling
 February 1953: suggested a triple
helix structure for DNA
11 Discovering the DNA Structure

James Watson & Francis Crick

 1953: Constructed the double-
helical model for DNA

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12 Discovering the DNA Structure
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DNA Structure

A double- stranded molecule with a

long chain of nucleotides

Nucleotide
Single building block of DNA
Deoxyribose sugar, phosphate
group, and nitrogenous base
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DNA Structure

Nitrogenous bases
Information-containing parts of
DNA
Purines: adenine (A) and
guanine (G)
Pyrimidines: cytosine (C) and
thymine (T)
Complementary base pairs
A <-> T
G <-> C
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DNA Structure

Sugar-Phosphate Backbone
Formed when the nucleotides
are joined into long chains
when strong attachments
called phosphodiester bonds
form between the
deoxyribose sugars and the
phosphates
Antiparallelism: opposing
orientation of the 2
nucleotide chains in a DNA
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How are DNA molecules wound so that

they fit inside the cell nuclei?

DNA PACKAGING
“To fit inside the cell nuclei, the DNA
molecules must fold so tightly.”
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DNA Packaging

Chromatin Condensed
DNA Nucleosome Chromatin loops
chromatin
loops
Chromosome

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DNA Packaging
1. DNA is wrapped around special protein molecules called
histones.
2. The combined loop of DNA and protein is called a
nucleosome.
3. Nucleosomes are packaged into a thread, which is
sometimes described as "beads on a string". Linkers tighten the
nucleosomes into fibers 30 nm in diameter. The end result is a
fiber known as chromatin.
4. Chromatin fiber is coiled. This fiber is then looped and coiled
yet again, leading finally to the familiar shapes known as
chromosomes.
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DNA Replication
20
DNA Replication

Why is it that the DNA must be replicated?

“DNA must be replicated so that the

information it holds can be maintained and
passed to future cell generations.”
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DNA Replication

Mode of DNA Replication:

1. Semiconservative
replication
- Replicated DNA would
consist of one “old” and
one “new” strand

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DNA Replication

Mode of DNA Replication

2. Conservative replication
- 2 newly created strands
are brought together & the
parental strands
reassociate

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DNA Replication

Mode of DNA Replication

3. Dispersive replication
- Each strand would consist
of both old and new DNA

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DNA Replication

DNA replication occurs

during S phase
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DNA Replication

OVERVIEW:
1. Parent DNA molecule
2. Parental strands unwind/unzip and separate at
several points.
3. Each parental strand serves as template for
complementary nucleotides to H-bond
4. Sugar-phosphate backbones of daughter strands
close
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3
5

2
4
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DNA Replication
SUMMARY:
1. Helicase binds to origin and separating strands.
2. Binding proteins keep strands apart.
3. Primase make a short stretch of RNA on the DNA template.
4. DNA polymerase adds DNA nucleotides to the RNA primer.
5. DNA polymerase proofreading activity checks and replaces incorrect
bases.
6. Leading strand synthesis continues in a 5’ to 3’ direction.
7. Lagging strand synthesis produces Okazaki fragments on the 5’ to 3’
template
8. DNA polymerase removes RNA primers and ligase seals sugar-
phosphate backbone
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Polymerase Chain Reaction (PCR)

DNA amplification
 DNA replication conducted
outside cells in a biotechnology
Polymerase Chain Reaction
(PCR)
 1st & best-known DNA
amplification technique
 Uses DNA polymerase to rapidly
replicate a specific DNA
sequence in a test tube Loading DNA samples for PCR
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Uses of PCR:

PCR has been used to amplify DNA from:

1. A cremated man, from skin cells left in his electric shaver, to
diagnose an inherited disease in his children.
2. A preserved quagga (relative of a zebra) and a marsupial wolf,
both extinct.
3. Microorganisms that cannot be cultured for study
4. Brain of a 7,000-year-old human mummy
5. Digestive tracts of carnivores, to reveal food web interactions
6. Roadkills and carcasses washed ashore, to identify locally
threatened species
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Uses of PCR:

7. Products illegally made from endangered species

8. Genetically altered bacteria that are released in field tests, to follow
their dispersion.
9. One cell of an 8-celled human embryo to detect a disease-related
genotype
10. Poached moose meat in hamburger
11. Remains in Jesse James’s grave, to make a positive identification.
12. Guts of genital crab lice on a rape victim, which matched the DNA of
the suspect
13. Fur from Snowball, a cat that linked a murder suspect to a crime
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PCR Requirements:
1. Primers:
 synthetic oligonucleotides (short
DNA strands) of a known
sequence of 15-30 bases
2. To keep the DNA strands
separated, processing must
be carried out at a relatively
high temperature
 Necessitates the use of DNA
polymerase isolated from
thermophilic bacteria: (a) Taq
polymerase from Thermus
aquaticus; and Vent polymerase
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32 PCR Requirements:

3. Thermal cycler
Machine that automatically
initiates the cyclic
temperature changes

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PCR Basic Steps

1. Denaturation
 Heating target DNA to 94◦C to separate it into 2 strands
2. Priming
 Oligonucleotide primers base-pair with the specific sites on the template
strands (between 50◦C and 65◦C)
3. Extension
 DNA polymerase is activated & begins to synthesize the DNA from a pool
of available nucleotides (72◦C)
34
Sequencing DNA

Frederick Sanger
 1977: invented a way to determine the base sequence of a
small piece of DNA (Sanger sequencing)
Sanger sequencing
 Generates a series of DNA fragments of identical sequence
that are complementary to the DNA sequence of interest
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Sanger Sequencing

 Uses an approach called “chain termination”

 Key principle: usage of dideoxynucleotide triphosphates (ddNTPs) as
DNA chain terminators
 Requirements:
Single-stranded DNA template
DNA primer
DNA polymerase
Radioactively/fluorescently labelled nucleotides
Modified nucleotides that terminate DNA strand elongation
36
Sanger Sequencing
Sequence of interest: TACGCAGTAC

Complementary seq.: ATGCGTCATG

Series of fragments: TGCGTCATG

GCGTCATG
CGTCATG
GTCATG
TCATG
CATG
ATG
TG
G
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Next Generation Sequencing

 Ability to sequence millions of small pieces at once that can

handle much larger DNA molecules much faster
 General approach is also called massively parallel DNA
sequencing
 A human genome can be sequenced in under a day
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Transcription
39 Transcription Copies the Information in
DNA

Human genes
 Encode 20,325 types of proteins
Proteins
 Consist of 1 or more long chains of amino acids
(polypeptides)
 Short sequence of amino acids is called peptide
 Bonds that join amino acids is called peptide bond
40 Protein Diversity in the Human Body
Protein Function

Actin, myosin, dystrophin Muscle contraction

Antibodies, antigens, cytokines Immunity

Carbohydrases, lipases, proteases, Digestion (digestive enzymes)

nucleases
Casein Milk protein

Collagen, elastin, fibrillin Connective tissue

Colony-stimulating factors, erythropoietin Blood cell formation

DNA and RNA polymerase DNA replication, gene expression

Ferritin Iron transport in blood

41 Protein Diversity in the Human Body
Protein Function

Fibrin, thrombin Blood clotting

Growth factors, kinases, cyclins Cell division

Hemoglobin, myoglobin Oxygen transport

Insulin, glucagon Control of blood glucose level

Keratin Hair structure

Tubulin, actin Cell movements

Tumor suppressors Cell cycle regulation

42 A cell uses 2 processes to manufacture
proteins using genetic instructions:

Transcription
 Synthesis of RNA molecule that is complementary to 1 strand
of the DNA double helix for a particular gene

Translation
 Uses the information in the RNA to manufacture a protein by
aligning & joining specified amino acids
43
RNA Structure
RNA
 Bridge between gene and protein
Coding strand
RNA polymerase
Ribonucleotide

RNA
Template strand

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DNA RNA
1. Usually double-stranded 1. Usually single-stranded
2. Thymine as base 2. Uracil as base
3. Deoxyribose as the sugar 3. Ribose as the sugar
4. Maintains protein- 4. Carries protein-encoding
encoding information information and controls
how information is used
5. Cannot function as an 5. Can function as an
enzyme enzyme
6. Persists 6. Transient
45 Three Major Types of RNA

1. Messenger RNA (mRNA)

 Carries information that specifies a particular protein
 Each 3 mRNA bases in a row form a genetic code word
(codon)
46 Three Major Types of RNA

The Genetic Code

47
Three Major Types of RNA

2. Ribosomal RNA (rRNA)

 Associate with certain
proteins to form a ribosome
 Some catalyze the formation
of the peptide bonds
between amino acids
(ribozyme)
 Some help align the
ribosome and mRNA
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Three Major Types of RNA

3. Transfer RNA (tRNA)

 Binds an mRNA codon at 1
end & a specific amino acid
at the other
Loop 3
 One loop of the tRNA has 3 Loop 1
bases in a row forming the
anticodon
 tRNA with a particular
anticodon strongly bonds to a
specific amino acid. Loop 2
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Three Major Types of RNA

Type of RNA Size (no. of Function

nucleotides)
Messenger RNA 500-4,500 Encodes amino acid sequence
(mRNA)
Ribosomal RNA 100-3,000 Associates with proteins to form
(rRNA) ribosomes, which structurally
support and catalyze protein
synthesis
Transfer RNA (tRNA) 75-80 Transports specific amino acids to
the ribosome for protein synthesis
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Steps of Transcription

1. Initiation
2. Elongation
3. Termination

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Setting the stage for transcription
to begin:
Transcription factors
 bind DNA at certain
sequences and initiates
transcription at specific sites
on the chromosomes
 TATA binding protein: 1st
transcription factor to bind

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Initiation
54
Elongation
55 To determine the sequence of RNA
bases transcribed from a gene:
Write the RNA bases that are complementary to the template
DNA strand.
Template DNA strand:
3’ CCTAGCTAC 5’
RNA complementary bases:
5’ GGAUCGAUG 3’
What will be the coding / nontemplate DNA sequence?
5’ GGATCGATG 3’
56
Translation
57
Translation

 Assembles a protein using the information in the mRNA

sequence
 Particular mRNA codons correspond to particular amino acids
 Genetic code – correspondence between the chemical
languages of mRNA and protein
 Takes place in the free ribosomes in the cytoplasm as well as on
the ribosomes that are embedded in the ER
58
Genetic Code

1. The code is triplet.

“If a codon consisted of 3
bases (any of the A, C, G, and
U), then the genetic code
could specify as many as 64
different amino acids.”

.: The minimum no. of bases in

a codon is three.
59 Genetic Code
Altering the DNA sequence by 1 or 2 bases produced
a different amino acid sequence.

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Genetic Code

2. The code does not overlap.

Example: mRNA sequence AUGCCCAAG
Nonoverlapping: AUG (methionine)
CCC (proline)
AAG (lysine)
61 Genetic Code

Overlapping:
AUGCCCAAG
AUG (methionine)
UGC (cysteine)
GCC (alanine)
CCC (proline)
CCA (proline)
CAA (glutamine)
AAG (lysine)
 62
Genetic Code

3. The code includes control.

AUG – signals “start”

UGA, UAA, and UAG – signify “stop”
63
Genetic Code
4. The code is the same in all species.

“All species use the same mRNA codons to specify the same amino
acids, and therefore the same genetic code.”

Universality of genetic code: All life evolved from a common

ancestor.

Exceptions: Few codons in mitochondria and in certain single-celled

eukaryotes (ciliated protozoa).
64 Three Major Types of RNA

The Genetic Code

65 Genetic Code

Synonymous codons – different codons that specify

the same amino acid

Nonsynonymous codons – encode different amino

acids
66
Building a Protein

Requirements:
mRNA
tRNA
Ribosomes
Energy-storing molecules (ATP & GTP)
Protein factors
67
Translation Steps:

1. Initiation
2. Elongation
3. Termination
4. Protein processing and
folding
 68 Initiation

mRNA leader sequence forms H

bonds with a short sequence of rRNA
in a small ribosomal subunit.

Start codon = AUG

Small ribosomal subunit + mRNA +

initiator tRNA
 Form the initiation complex
69
Elongation

Filling of A site by a second

tRNA
70
Elongation

Formation of peptide bonds

71
Elongation

Discharge of tRNA 1 at E site

(translocation)
Enzyme-directed shifting
of the ribosome to the
next position on the mRNA
strand
72
Elongation

tRNA 2 shifts into P site

tRNA 3 enters ribosome at
A site
73
Elongation

Formation of peptide bond

74 Elongation

Discharge of tRNA 2
2nd translocation
tRNA 4 enters ribosome
75 Elongation
 76
Termination
Presence of at least one
special codon occurring
just after the codon for the
last amino acid

Termination codons/stop
codons/nonsense codons:
UAA, UAG, UGA

Message: STOP HERE

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77 Protein Processing

Proteins fold into 1 or more 3D shapes or

conformations:
1. Primary (1º) structure
2. Secondary (2º) structure
3. Tertiary (3º) structure
4. Quaternary (4º) structure
78
Primary (1º) Structure

Amino acid sequence of

a polypeptide chain

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Secondary (2º) Structure

Chemical attractions between

amino acids that are close
together in a 1º structure fold
the polypeptide chain into its
secondary (2º) structure

2 common secondary
structures:
1. Alpha helix
2. Beta-pleated sheet
80
Tertiary (3º) Structure

3D form, secondary structure

that are wound due to the
attraction and repulsion of
more widely separated amino
acids in response to water
molecules
81
Quaternary (4º) Structure

Protein consist of more than

one polypeptide

Example:
Hemoglobin – 4 polypeptide
chains
82
Protein Folding

Proteins begin to fold within a minute after the amino acid chain
winds away from the ribosome

1. Chaperone proteins – stabilize partially folded regions in their

correct form & prevent a protein from getting “stuck” in an
intermediate form, which would affect its function
2. Ubiquitin – straighten & refold the misfolded protein
3. Proteasome – degrade the protein into amino acids
83
Protein Folding

Proteins misfold in 2 ways:

1. Mutation
May change the amino acid
sequence
2. Having more than 1 conformation
2 forms of the same protein have
identical amino acid sequences, but
fold differently
Example: prion
84
Prion Diseases

 Scrapie = sheep
Denotation: infected sheep rub
against things to scratch themselves
Brains of the infected sheep: riddled
with holes
More than 85 animal species
develop similar disorders
 Creutzfeldt-Jakob disease, fatal familial
insomnia, and Gerstmann-Straüssler-
Scheinker disease = humans
85
Prion Diseases
Kuru (means “to shake”)
 1st prion disease recognized in humans
 Affected the native Fore people who lived in
remote mountains of Papua New Guinea
 Wobbly legs, trembling, & whole body
shaking
 Uncontrollable laughter (“laughing disease”)
 Speech slurred, thinking slowed, & the person
became unable to walk or eat
 Death came within a year
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 What is the complementary
11 DNA sequence given the
template DNA strand:
TACCGCTTAAACCTCCATACT
88
REFERENCES

 Klug, W. S. & Cummings, M. R. (2002). Essentials of Genetics 4th edition.

Upper Saddle River, NJ: Prentice-Hall, Inc.
 Lewis, R. (2015). Human genetics: concepts and applications 11th edition.
New York:McGraw-Hill Education
 Sundara Rajan, S. (2000). Cytogenetics. New Delhi: Anmol Publications Pvt.
Ltd.