Genetic Material,

Genetic Code and

Protein Synthesis

BY ANANYA ARAVIND

http://www.free-powerpoint-templates-design.com
IMPORTANT TERMS
1. Codon
2. Genetic code
3. Genome
4. Replication
5. Recombination
6. Deoxyribonuclease
7. DNA ligase
8. Polymerase
9. Central dogma
10.Transcription
11.Sense strand
12.Transformation
13.Translation
 Codon a sequence of three nucleotide bases (in mRNA) that codes for an amino acid or the the
initiation or termination of a polypeptide chain

 Genetic code the order of base pairs along the DNA molecule that controls the kind and order of
amino acids found in the proteins of an organism

 Genome a complete set of genetic material

 Replication production of a strand of DNA from the original

 Recombination formation in daughter cells of gene combinations not present in either parent

 Deoxyribonuclease an enzyme that catalyzes the hydrolysis of DNA to nucleotides

 DNA ligase a specific enzyme that joins fragments of replicated stands together

 Polymerase an enzyme that links nucleotides together to form polynucleotide chains

 Central dogma the process by which the instructions in DNA are converted into a functional product

 Transcription the process in which a complementary single-stranded mRNA is synthesized from one of the DNA
strand

 Sense strand the DNA strand which serves as template for transcription

 Transformation the phenomenon by which DNA isolated from one type of cell when introduced into another type
is able to bestow some of the properties of the former to the latter

 Translation the process in which genetic information in mRNA directs the order of assembly of the specific
amino acids during protein synthesis
Properties of Genes
It should be able too
01 replicate

It should be chemically and

02 structurally be stable

It should be able to express

03 itself in the form of
Mendelian Characters
04 It should be able to mutate
Evidences for DNA as genetic material

HERSHAY
GRIFFITH’S AVERY,MACLEOD
AND MCCARTY’S AND
EXPERIMENT
EXPERIMENT CHASE’S
EXPERIMENT
Griffith’s Bacterial
Transformation Experiments
 Two strains of D.pneumoniae observed
 Smooth (S) type
 Rough (R) type

S type R type
Large polysaccharide No
capsule around the cell polysaccharide
Has a glistening slime layer
appearance Has an irregular
Virulent- can cause appearance
pneumonia Non virulent
AVERY, MACLEOD AND MCCARTY’S EXPERIMENT
 It suggested that DNA and not proteins
is the genetic material

 They found that the DNA isolated from

heat killed S-cells when added to
R-cells changed their surface
character from rough to smooth and
also made them pathogenic.

 When the extract was treated with

DNAase (an enzyme which destroys
DNA) the transforming ability was lost.

 Proteases (enzymes which destroy

proteins) however did not affect the
transforming ability
 HERSHEY AND CHASE EXPERIMENT
 They conducted experiments on the bacteriophages that attack
bacteria.

 The bacteriophage‘s injecting genetic material that takes over the

bacterial metabolic machinery forces to produce new bacteriophages.

 They separately labelled the protein and DNA component of the

bacteriophages with specific radioactive tracers and then followed the
components through the phages' life cycles

 Almost all proteins contain sulphur an atom not found in DNA

whereas all DNA molecules contain phosphorus an atom not found in
proteins .
 Thus bacteriophages parasitizing bacteria grown
in the presence of radioactive sulphur had
labelled proteins and the bacteriophages
parasitizing bacteria grown in the presence of
radioactive Phosphorus had labelled DNA

 It enables to distinguish between DNA and

proteins of the phage

 They combined each strain with non radioactive

bacteria and allowed bacteriophages to attach
and inject their genetic material .

 After agitation it was observed that only 32P

Radioactive was found associated with
bacterial cells and 35S was only in the
surrounding medium and not in the bacterial cell
.

 Even progeny carried label only with 32P not

with 35S

 This clearly indicate that only DNA and not

protein coat enters the bacterial cell
BASICS
DNA is composed of three different chemical substances
• A five carbon (pentose) sugar deoxyribose
• A phosphoric acid group
• 4 nitrogenous bases
Purine- Adenine and Guanine
Pyrimidine- Cytosine and Thymine
(Uracil in RNA)

CHARGAFF’S RULE
 The total molar amount of adenine in any species
specimen of DNA is always equal to that of thymine
 Thus the ratio A/T is always 1.
 Similarly the amount of cytosine is equal to that of
guanine.
WHICH IS THE GENETIC MATERIAL?
 DNA
• In DNA the true stands be complementary if
separated by heating come together when
appropriate conditions are provided
• DNA is less reactive and structurally more stable
when compared to RNA
• The presence of timing at the place of uracil also
confers additional stability to DNA
• DNA is dependent on RNA for synthesis of proteins
• DNA being more stable is preferred for storage of
genetic information
RNA
• 2-OH group present at every nucleotide in RNA
is a reactive group and makes RNA labile and
easily degradable.
• RNA is reactive due to its catalytic property
• RNA being unstable mutate at a faster rate
• RNA can directly code for the synthesis of
proteins and can easily Express the characters
• RNA is better for transmission of genetic
information
Using Analysis of X Ray diffraction pattern of DNA,
the important features of DNA as revealed by
Franklin and Wilkins are as follows:

 It has a helical structure

 It is long and thin with uniform diameter of
20ÅThe distance between successive
nucleotides is 3.4Å
 The helix makes 1 complete turn every 34Å
long its length
 There are 10 nucleotides per turn of helix
Double helix model
as proposed by Watson and Crick
 In each DNA molecule there are two long and parallel
polynucleotide chains helically coiled around the same axis
 The right-handed helix are held together by their bases which are
paired together by covalent hydrogen bonds.
 Both the chains of a DNA molecule are thus complementary to
each other
 There are 10 bases for turn of double helix
 One turn of helix is as such completed in 34Å
 The two helix are coiled in such a way that they cannot be
separated without unwinding
 The backbone of the helix is a chain of sugars and phosphate
alternating with each other
 The two chains of a helix are of opposite polarity (antiparallel)
 If one chain runs in 3 - 5 direction the other will run in 5 - 3
direction
 The width of the helix is 20Å
 Length of DNA is characteristic of an organism
 STRUCTURE OF RNA
RIBOSOMAL RNA (rRNA)
FEATURES
.
 It is normally single
stranded structure
 Uracil replaces
thymine in the RNA
MESSENGER RNA (mRNA)
molecule
 Adenine pairs with
uracil in RNA
 In RNA the pentose
sugar is ribose TRANSFER RNA (tRNA)
instead of
deoxyribose
 Ribosomal RNA
 It is the most abundant of all types of
RNA
 Makes up about 80 percentage of the
total RNA of a cell
 It is single standard and most stable
form of RNA
 That has the highest molecular weight
 The unpaired bases in rRNA molecule
binds mRNA and tRNA to ribosomes
(Mg++ linkage)
 Messenger RNA
 It is most heterogeneous in size and stability
 it forms a template for protein synthesis
 A particular region of the DNA molecule is
copied during the synthesis of RNA by
replacing thymine with uracil and the sugar is
ribose instead of deoxyribose
 The size of mRNA depends on the size of
proteins is codes for.
 mRNA carries coded message from DNA to
ribosomes
 It directs amino acid sequence in protein
synthesis
 mRNA is short lived
 Transfer RNA
• It acts as vehicles that pickup amino acid
scattered throughout the cytoplasm and
transport them to specific codons of mRNA
molecules on ribosome (i.e they are assist in
protein synthesis)
• Hence it is also called an adaptor molecule
• It is held in a clover leaf shape by hydrogen
bonds
• The matching of anticodon with amino acid is
done by an enzyme-amino acid activating
enzyme called amino acyl-tRNA synthetase
• Each type of t RNA molecule reflects a
complementarity in base composition and
sequence of a particular gene in the DNA
The following three hypothesis of
DNA replication have been proposed

Conservation replication
01
Original double helix serve as a template and an entirely new double-
stranded molecule is synthesized .

Dispersive replication
02 The original molecule is broken into fragments; each fragments serves as
a template for the synthesis of complementary
05
fragments and finally two
new molecules are formed which consists of both old and new fragments .

Semi-conservative replication
03 The separated single strands act as templates for the synthesis of new strands
each daughter double helix carries
06 one polynucleotide strand from the parent
molecule and one newly assembled strand
Meselson-Stahl Experiment
 The Meselson-Stahl experiment was an experiment to
prove that DNA replication was semiconservative.

 Semiconservative replication means that when the double

stranded DNA helix was replicated, it would produce two
copies which contained one of the original strands, and one
entirely new copy.

 The deciphering of the structure of DNA by Watson and

Crick in 1953 suggested that the semiconservative model
was correct

 Nitrogen is a major constituent of DNA, the genetic material

of all cells.

 It is commonly found in the 14N isotope, but it can also be

found in the heavier 15N isotope.
 E. coli were grown for several generations in a medium
with 15N.The DNA of the resuting cells had a higher
density (was heavier).

 E. coli cells with only 15N in their DNA were put back into
a 14N medium and were allowed to divide only once.

 DNA was then extracted from a cell and was compared

to DNA from 14N DNA and 15N DNA.

 It was found to have exactly an intermediate density. This

supported the idea of semiconservative replication.

 The DNA was intermediate in density because it had one

15N DNA strand and one14N DNA strand. The 15N
strand was one of the original strands in the original cell.
The 14N strand was a newly synthesized strand.
DNA Replication
Origin of Replication

Replication begins at the initiation site

called origin
Prokaryote - one origin
Eukaryote - several origins
Helicase (rep protein)

At the origin, enzymes relax facilitated

by DNA gyrase (topoisomerases)
Unwinding facilitated by Helicase
Replication fork

The strands are pulled apart forming a

Y shaped structure
Primer

 Short stretch of RNA formed on the DNA template

 It produces 3'OH end on the sequence of ribonucleotides
to which deoxyribonucleotides are added
• Primase
An enzyme which polymerises RNA building blocks into
primer
 DNA polymerase

The newly synthesized complementary nucleotides are paired with the

the existing nucleotides on the the parent strands and covalently
bonded together by a complex of enzymes called DNA polymerases

Energy used by the DNA polymerase for the covalent bonding comes
from the newly synthesized nucleotides (triphosphate)

 Finally the nucleotides are in position, gaps are sealed by another

enzyme DNA ligase
 Adding nucleotides

 As two strands of DNA helix have an antiparallel

orientation they require different treatment by the DNA
polymerase
 This enzyme catalysed the addition of nucleotide to this you
stands in the 5'-3' direction as it can only add nucleotides to the
3' carbon position
Leading Strand

 Thus for one of the parental stand, there is always a 3

position at the replication fork and therefore its new
complementary strand can be synthesized continuously in
the 5’-3’ direction
Lagging Strand

 For the other parents stand however there is always a 5’

carbon at the replication fork so its new strand of
nucleotides must be synthesized backward in
pieces(Okazaki fragments)
 These short pieces are later joined together by the
enzyme DNA ligase after replacing the RNA primer with
DNA
CENTRAL DOGMA
PROTEIN SYNTHESIS
Transcription
TRANSCRIPTION
It is the first process in the
overall information transfer
from DNA into protein
structure
 As a portion of a DNA molecule uncoils, along the exposed sequence of bases of one of the two strands of DNA
a complementary strand of mRNA is synthesized from DNA strand template sense strand

 DNA strands
• strand – template – sense strand
• Strand – coding strand – antisense strand

 The reaction is catalysed by the enzyme RNA polymerase

 Prokaryotes - a single RNA polymer - all three types of RNA
 Eukaryotes
• RNA polymerase I (nucleolus) rRNA
• RNA polymerase II (nucleoplasm) mRNA
• RNA polymerase III(nucleoplasm) tRNA
Two strands of the DNA have
opposite polarity .

RNA polymerase catalyzes the

polymerization in only one direction
5’ – 3’ direction.

The strand that has the polarity 3’ –

5’ acts as a template.

The others stand (coding strand)

does not code for anything .
 Structural gene
• Polycistronic (Prokaryotes)
• Monocistronic (Eukaryotes)

 Monocistronic (Eukaryotes)
Intron - noncoding segment
of a gene
Exon - that portion of DNA
which codes for the
final RNA
Transcription unit in DNA
Promoter
 located towards 5’ end of the coding strand
 Defines beginning of the prescription

Structural gene
 DNA sequence that provides binding site for RNA polymerase

Terminator
 located towards 3’ end of the coding strand
 Defines end of the transcription
Trancription in Eukaryotes

• Splicing
Introns are removed and exons are
joint in a defined order.

• Capping
Unusual nucleotide (methyl
guanosine triphosphate) is added to the 5'-
end hnRNA (heterogenous nuclear).

• Tailing
Adenylate Residues(200-300) was
added at 3'-endin a template independent
manner
Translation
–
Nature of
GENETIC CODE
1.Triplet
2.Universal
3.Non-overlapping
4.Degenerate
5.Commaless
6.Non-ambiguous
 1. Triplet
 The minimum
requirement is a triplet
code
 Total 64 amino acids
present
2. Universal

 Universal genetic code means that all known living system use nucleic
acids and the same three base codons to direct the synthesis of proteins
from amino acids
Exception
In mitochondrial codons and in some protozoans
3.Non- overlapping
 It means that the same
letter is not used for two
different codons
4. Degenerate
 It means that more than
one triplet sequence
could code for a specific
amino acid
5. Commaless
 No punctuations are needed
between any two codes.

 After one amino acid is coded,

the second one will be
automatically coded by the
next three nitrogen bases till
the stop codon occurs.
6. Non-ambiguous
I will always
encode Alanine

 It means that a particular

codon will always code for
the same amino acid
whenever it is found

 Exception

GUG – Valine
– Start codon
Chain initiation codon
AUG (Methionine)
GUG (Valine)

 When occured between two ends of cystone they code

for the amino acids
 But when they are used as initiating codon they do not
code for amino acid but initiate the synthesis of
polypeptide

Chain termination codon (nonsense codon)

UAA
UAG(Tyrosine)
UGA(Tryptophan)

 They do not code for any of the 20 essential amino

acids
 When any one of them occurs immediately before the
initiation triplet it causes the release of polypeptide
chain from the ribosome
DNA Packaging
–
Nucleoid
In prokaryotes, DNA is held
with some proteins in a region
termed as nucleoid
The DNA in nucleoid is
organised in large groups held
by proteins.
Chromosomes
 One long coiled or
folded DNA molecule
which is associated with
histone protein on
nucleoprotein
Nucleosome

 Units which when

repeated to form
chromatin fibre
resembling beads on
a string
Histones
 Set of positive charged basic protein
 Rich in basic amino acid Residue lysing
and arginine
 Organised to form a unit of eight
molecules called histone octamer
 Octomer of proteins consists of two
molecules each of the four different
histones tetramers
 These histones are H2A, H2B, H3, and
H4
 The core particles are linked by the
DNA which is associated with only one
type of histone H1
 EUCHROMATIN HETEROCHROMATIN
The region of chromatin which is The region of chromatin which is
loosely packed more densely packed

Stain light Stain dark

Transcriptionally active chromatin Transcriptionally inactive chromatin

 First level (10 nm)
• The nucleosome consists of a spiral of DNA
wrapped around an octomer of histone
molecules forming core particle
• The particles ( histones) are linked by DNA
which in turn is associated with only one type of
histone H1

 Second level (30 nm)

• 10nm fibre of nucleosomes get coil upon itself to
form 30 nm wide helix with five or six
nucleosomes per helix.
• In this helix, successive turns come close
together and this 30nm structure is called
solenoid .
• H1 protein molecules aggregate by crosslinking
to form polymers and thus help in folding of
10nm fibre into 30 nm solenoid.

 Final level
• Condensation of these fibres into metaphase
chromosomes where the DNA appears to loop
out from a scaffold of proteins
Thank you