CHAPTER#20 Chromosomes & DNA

Chromosomes:
Chromosomes are thread like structures that appear inside the nucleus at the time of cell division.

Discovery:
They were first observed by Walther Fleming in 1882 during study of salamander larvae.

Organism No. Of chromosomes

Mosquito 6
Honeybee 32
Corn 20
Sugarcane 80
Frog 26
Mouse 40

Function of Chromosomes:
They determine how a person’s body develops and functions.

Structure:
Typically a chromosome is made up of
chromatids (two replicas), centromere
(primary constriction) and a secondary
constriction.

Karyotype:

The particular array of chromosomes that an individual possesses is called its Karyotype.

Chromosomes are different regarding following ways:

 Size
 Staining properties
 Location of centromere
 Length of chromatids

Types of chromosomes:
The chromosomes arc usually classified on the basis of position of centromere.
 Telocentric chromosomes Acrocentric chromosomes
• The chromosome in which centromere is located • The chromosome in which
at one end and have only one arm is called centromere is located far away from the
telocentric chromosome. center and have a small arm and other
large arm is called acrocentric
chromosome.
• It has a single arm. • It has a very short and a very long arm.
• It gives i shape to chromosome. • It gives J shape to chromosome.

Metacentric chromosome Sub metacentric chromosome

• The chromosome in which centromere is • The chromosome in which
located at center and have two equal arms is centromere is not located at center
called metacentric chromosome. and have two unequal arms is
called metacentric chromosome.

• The two arms have equal length. • The two arms have unequal length.
• It gives V shape to chromosome. • It gives L shape to chromosome.
 Composition of chromosomes
Chromosomes are composed of DNA and protein. Mostly chromosomes are about:
• 40% DNA
• 60% protein
 A human chromosome contains about 140 million (1.4 x 108) nucleotides in its DNA.
 If the strand of DNA from a single chromosome were laid out in a straight line, it would
be about 5 centimeter long.
 The amount of information one chromosome contains is equal to:
 About 280 printed books.
 Each book contains 1000 pages.
 Each page had about 500 words on it.
Nucleosomes:
Every 200 nucleotides, the DNA duplex are coiled around a core of eight histone proteins
forming a complex known as a nucleosome.

Histones are positively charged proteins.

• Positive charge is due to an abundance of the basic amino acids:
 Arginine
&
 Lysine
• Nucleosomes are strongly attracted to the negatively charged phosphate groups of
the DNA.

Heterochromatin Euchromatin
• Highly condensed portions of the • Chromatin that is condensed only during
chromatin are called hetero-chromatin. cell division is called euchromatin.
• Heterochromatin is only present in • Euchromatin is present in both eukaryotes
eukaryotes. and prokaryotes.
• Heterochromatin is inactive. • Euchroamtin is highly active.
• Heterochromatin is found at the periphery of • Euchromatin is found in the inner body of
nucleus. the nucleus.
• It never exposes itself. • It is always present in open configuration.

CHROMOSOMAL THEORY OF INHERITANCE

The chromosomal theory of inheritance is the idea that “genes, the units of inheritance,
are found on the chromosomes, so chromosomes act as carriers of heredity”.
 Emergence or origin of chromosomal theory of inheritance is linked with discovery of
chromosomes which were first observed by Walther Fleming.
 The term “chromosome” was proposed by Waldeyer, which literally means colored bodies.
 First time the relationship of heredity units with chromosomes was put forward by Karl
Correns.
 The actual credit of this theory goes to both Walter Sutton and Theodor Boveri.
 In 1902, these scientists recognized independently that the behavior of Mendel’s factors
(genes) is parallel to the behavior of chromosomes at meiosis.
 If Mendel’s model is correct, then these two gametes must make equal hereditary
contributions. Sperm, however, contains little cytoplasm and during fertilization it only
contributes nucleus to the zygote.
 Therefore, the hereditary units must reside within the nucleus of the gametes.
 Genes would be present in chromosomes.
 Many investigators of that time pointed out a serious objection on Sutton’s theory.
 After some years the objection was cleared after the discovery of linkage by the historical
experimentation of T. H. Morgan in 1910 on Drosophila.
Parallel behavior of genes and chromosomes during meiosis.
Behavior of chromosomes Behavior of genes
1- Diploid cells (before meiosis) have two 1- According to the Mendel, diploid cells
copies of each chromosome have two copies of each gene (pair of
(homologous pairs) while gametes (after alleles) while gametes have only one.
meiosis) have only one. e.g. In pea plant, diploid cells have pairs of
E.g. In pea plant diploid cells have 7 pairs alleles for each gene like Rr, Yy, and Tt,
of homologous chromosomes while while gametes have single R or r, Y or y
gametes have single 7 chromosomes. and T or t
2- Homologous pairs of chromosomes 2- According to the Mendel, pair of gene
segregate during meiosis. for each trait also segregates from each
other during meiosis. e.g. Rr
3- During meiosis, each pair of 3- According to Mendel, alleles of one gene
homologous chromosomes orients on pair also assort independently to the
the metaphase plate independently of alleles of other gene pair during meiosis.
any other pair so that in anaphase each e.g. RrYy genotype as a result of
pair assorts independently of the other. independent assortment can form four
type of gametes i.eRY, Ry, rY, and ry.

DNA AS HEREDITARY MATERIAL

Work of Frederick Griffith
 Frederick Griffith was a British microbiologist who provided first evidence about
hereditary nature of DNA.
 He used Streptococcus pneumonia bacteria.
There are two types of S. pneumonia.

S-form R-form
S-form is virulent. R-form is non-virulent.
It contains polysaccharide coat It lacks an enzyme needed to
necessary for virulence. manufacture polysaccharide coat.
 He performed following experiments.
 When he infected mice with virulent strain of S. pneumonia S-form, it died of blood
poisoning.
 When he infected similar mice with mutant strain of S. pneumonae that lacked the
virulent strains polysaccharides coat R form, it did not cause the death.
 He injected dead bacteria of S-virulent strain into the mice, the mice remained perfectly
healthy.
 As a control, he injected mice with a mixture containing dead S bacteria of virulent strain
and live coatless R bacteria although each of them did not harm mice separated but their
mixture caused death of mice. In blood of dead mice live S bacteria were found.
Transformation
“It is the transfer of genetic material from one cell to another altering genetic makeup of
the recipient cell.”
From these experiments he concluded that some information specifying the polysaccharide
coat had passed from the dead, virulent S bacteria to the live, coatless R bacteria in the
mixture, permanently transforming the coatless R bacteria into the virulent S variety.

Work of Avery, Macleod and McCarty

They discovered agent responsible for transforming streptococcus. They performed
following experiments.
 They prepared mixture of dead S streptococcus and live R streptococcus and removed
much of the protein (99.98%) by applying protein digesting enzyme.
Transforming activity was not reduced.
 They removed much of RNA by applying RNA digesting enzyme.
Transforming activity was still present.
 They removed DNA by applying DNAase. At that time transforming activity was lost.
Work of Hershey and Chase
They performed experiment with bacteriophages (T2) supporting Avery’s conclusion.
(i) In an experiment, they labeled viruses with radioisotope 32P being incorporated into
newly synthesized DNA of growing phage.
(ii) In other experiment, they labeled viruses with radioisotope 35S being incorporated into
the amino acids of newly synthesized protein coats.
After labeled viruses were permitted to infect bacteria, bacterial cells were agitated
violently in a blender to remove the protein coats of the infecting viruses from the surface
of bacteria. This procedure removes nearly the entire 35S label from the bacteria.
However, 32P label had transferred to the interior of the bacteria and was found in viruses
subsequently released for the infected bacteria. Hence the hereditary information injected
into the bacteria that specified the new generation of viruses was DNA and not protein.
 Chemical Nature Of DNA
Discovery of DNA:
F. Miescher, discovered DNA in 1869.
Miescher extracted a white substance from the nuclei of
 Human cells
&
 Fish sperm cell.
He called this substance “nuclein” because it seemed to be specifically associated with the
nucleus.
Since nuclein was acidic, later on, it came to be known as nucleic acid.
In 1920’s, the basic structure of nucleic acids was determined by the P.A. Levene.
Main components of DNA:
 Phosphate (PO4-3) groups
 Five carbon sugar ( Ribose)
 Nitrogen containing bases called purines and pyrimidines.

Purines = (Adenine, A, and guanine, G)

Pyrimidines = (Thymine, T and cytosine, C, RNA contains uracil, U)

Structure: of nucleotide:
In a typical nucleotide:
 Nitrogenous base is attached to carbon number 1 of a pentose sugar.
 Phosphate group is attached to carbon number 5 of the sugar.
 Hydroxyl (-OH-1) group is attached to the 3 carbon atom.
Phosphodiester bond:
Bond b/w two nucleotides is called Phosphodiester bond.
Formation of Phosphodiester bond:
 The reaction between phosphate group of one nucleotide and hydroxyl group of another
nucleotide is a dehydration synthesis, eliminating a water molecule.
 The linkage is called a phosphodiester bond because the phosphate group is now linked to
the two sugars by means of a pair of ester (C-O-P-O- C) bonds.
 DOUBLE HELICAL STRUCTURE OF DNA (WATSON-CRICK’S MODEL)

Important features of the model are as follows:

 A DNA molecule has two un-branched complementary strands which are spirally coiled
and form DNA duplex. Both strands are antiparallel.
 Nitrogen containing bases called purine and pyrimidine.

Purine Pyrimidine
They are double ringed nitrogenous bases. They are single ringed nitrogenous bases.
It includes adenine (A) and guanine (G). It includes cytosine (C, thiamine (T) and
uracil (U).
They are larger in size. They are smaller in size.

 Two hydrogen bonds form between A & T.

 Three hydrogen bonds form between G & C.
 One turn of the spiral has 10 nucleotides on
each strand.
 Each turn occupies a distance about 3.4 nm.
 Distance between adjacent nucleotides is 0.34 nm.
 The back bone of DNA chain is built up of deoxyribose
and phosphoric acid.
 Diameter of DNA is 2 nm.
 Two chains are held together by hydrogen bonds.

Services of Erwin Chargaff:

Erwin Chargaff showed that:
 The amount of adenine in DNA always equals the amount of thymine.
 The amount of guanine always equals the amount of Cytosine.
Services of Rosalind Franklin:
Rosalind Franklin prepared this X-ray diffraction pattern of DNA in the laboratory.
Process:
 In this analysis a molecule is bombarded with a beam of X-rays.
 When individual rays encounter atoms their path is bent or diffracted and the
diffraction pattern is recorded on the photographic film.
 When carefully analyzed this pattern gives three dimensional structure of a molecule.
Conclusion:
The diffraction pattern prepared suggested that:
 DNA molecule had a shape of a helix.
 Diameter DNA of 2nm.
 One complete helical turn is of 3.4 nm.

DNA REPLICATION
Features HYPOTHESIS 1 HYPOTHESIS 2 HYPOTHESIS 3
Name Conservative Semi-conservative Dispersive
Statement When both parental "Two strands of a DNA "Breaks the DNA
strands stay together molecule separate backbone every 10
after replication is during replication, nucleotides or so,
called conservative Each strand acts as a untwists the molecule,
model. template for the & attaches the old
synthesis of new strand. strand to the end of
newly synthesized one.
Unwinding of Done Done Not done
2 strands
After One duplex old + All One old + One new Each duplex is mixture
replication duplex new strand in each duplex of old and new DNA
 Meselson and Stahl Experiment
 They used DNA of E. coli bacteria.
 They proved that DNA replication is done by
semi-conservative way.

PROCEDURE:
First media:
 They growth bacteria in a medium containing heavy
isotope of nitrogen N-15.
 When grown on medium containing heavy N-15
the bacteria took up N-15
nitrogen and used it to synthesize new biological
molecules, including DNA.
Second media:
 Then they transferred bacteria from N-15 to N-14
medium and collected DNA at different intervals.
Centrifugation:
 They dissolved the different samples of collected
DNA in cesium chloride and spun it at very high
speed in an ultra-centrifuge.
 Ultracentrifuge gradient of CsCl2 i.e. Cs+2 moves towards bottom of test tube & Cl-1
moves on top of tube.
 As Cesium is heavy and Chloride is light, so their densities match with N-15 & N-14
respectively.
Consequently,
 N-15 moves to bottom of test tube.
 N-14 moves on top.
 Hybrid strand suspends in middle of test tube.
Results:
Zero round of replication:
 The DNA collected immediately after transfer (sample at 0 minute) was all dense i.e. N-
15.
First round of replication F1:
 The DNA collected 20 minutes after transfer was intermediate between N-14 & N-15.
Second round of replication F2:
 The DNA collected 40 minutes after transfer was of two classes.
i. One was intermediate between N-14 & N-15.
ii. One was light strand N-14 only.
 THE REPLICATION PROCESS
Definition:
Formation of DNA from DNA is called Replication of DNA.

Replication origin:
The DNA replication begins at one or more sites on the DNA molecule, where there is a
specific sequence of nucleotides. This specific site is called replication origin.
Enzymes/ proteins involved:
Helicase:
It opens double helix of DNA by breaking hydrogen bonds.
SSBPS:
They prevent recoiling of DNA after unwinding.
Primase:
It constructs an RNA primer i.e. a sequence of about
10 RNA nucleotides.
DNA polymerase:
It catalyzes the addition of d-nucleotides to the complementary growing DNA strands.
Properties of DNA polymerase:
 It can add nucleotides only to a chain of nucleotides that is already paired with the
parent strands.
 They are of three types I, II and Ill in bacteria.
 It can add nucleotides to the 3’ end of a DNA strand so replication always proceeds
from 5’ 3’direction on a growing DNA strand.
 The true E. coli replicating enzyme is DNA polymerase III which is 10 times larger.
 DNA polymerase cannot initiate synthesis on its own, it needs primer.
 This enzyme is a dimer and catalyzes replication of one DNA strand.
DNA Ligase:
It joins the Okazaki fragments formed in lagging strand of DNA replication.

Types of DNA polymerase

DNA polymerase I DNA polymerase II DNA polymerase III
 It plays supportive role.  It proof reads the  It involves the
 Converts RNA complementary nucleotides addition of d-ribonucleotides
nucleotides of primer into by moving from 3 5. in the direction of 5 3 with the
DNA nucleotides.  It plays role in DNA speed of 1000 nucleotides per
repair. second.

Mechanism:
Following steps are involved during DNA replication.
i) Helicase opens double helix of DNA and SSBPs prevent recoiling.
ii) Primase adds primer complentary to DNA strand.
iii) DNA polymerase III recognizes primer and constructs new strand in 5’ 3’.
iv) Leading & lagging strands are formed during DNA replication.

Leading strand Lagging strand

• The strand of DNA that is assembled • The strand of DNA that is assembled away
toward the replication fork is called leading from the replication fork is called lagging
strand. strand.
• Leading strand exhibits continuous • Lagging strand exhibits discontinuous
replication. replication.
• Okazaki fragment cannot be formed in • Okazaki fragment can be formed only in
leading strand synthesis. leading strand synthesis.
• DNA replication on the leading strand needs • DNA-ligase is required for lagging strand
to be primed only once. synthesis.

v) When the DNA polymerase III reaches the 5 end of lagging strand, DNA ligase
connects these okazaki fragments.
vi) The DNA molecule is further unwound, new RNA primers are constructed & DNA
polymerase III then jumps ahead 1000-2000 nucleotides (towards the replication
fork) to construct another fragment.

Okazaki fragments:
The short segments formed in lagging strand
due to its discontinuous synthesis during DNA
replication are called Okazaki fragments.
 Each segment is synthesized from 3 5.
Okazaki fragments are:
 100-200 nucleotides long in eukaryotes.
  1000-2000 nucleotides long in prokaryotes.

What is Gene?
Each gene produces its effect by controlling the synthesis of a particular enzyme.
This is called one-gene one-enzyme hypothesis. Since each enzyme is formed of
polypeptides so this hypothesis may be known as one-gene one-polypeptide hypothesis.
Work of Garrod and Bateson:
Archibald Garrod and William Bateson concluded in 1902 that certain metabolic heredity
diseases were more prevalent in particular families.
Alkaptonuria:
 In Alkaptonuria the patients produced urine that contained homogentisic acid.
 This substance oxidized rapidly when exposed to air turning the urine black.
 In normal individuals, hormogentisic acid is broken down into simpler substances.
Conclusion:
 They concluded that patients suffering from Alkaptonuria lacked the enzyme necessary to
catalyze this breakdown.
 They speculated that lack of enzyme is due to mutation in any gene.
CENTRIBUTION OF BEADLE AND TATUM:
Neurospora (Wild type):
They selected wild type of neurospora ( a fungus known as red bread mold for their experiments,
which could grow on minimal medium.
Minimal Medium:
It was the medium that provided a few substances for the growth of the mold.
It contained only:
 Sugar
 Ammonia
 Salts
 Few vitamins
  Water
Neurospora can synthesize all of the other
substances on its own because it contains
normal enzymes,

Exposing of spores to X-rays:

 Beadle and Tatum exposed spores of Neurospora to X-rays.
 They were expecting that DNA in some of these spores would get damage in their ability
to make compounds needed for normal growth.

Mutation Mutant
DNA changes of this kind are called The organisms that have undergone such
mutations changes are called mutants.

Growth of Irradiated Spores:

 They allowed the progeny of the irradiated spores to grow in a complete medium.
 Compelte medium contains all of the nutrients necessary for growth,
 Any growth deficient mutants resulting from the irradiation could take up the required
substance from the complete medium and would be kept alive.
DETERMINATION OF MUTANTS:
 To determine mutation in any of spore, Beadle and Tatum placed subcultures of
individual fungal cells again on minimal medium.
 Cells that had lost the ability to make other compounds necessary for growth would not
survive on such a medium.
 They added one by one various chemicals to the minimal medium in an attempt to find
out which chemical would enable a given mutant strain to grow.
Conclusion:
Among various chemicals they tried, the addition of arginine, permitted several mutant strains,
dubbed (named as) arg mutants to grow.

Mutation in Chromosomes:
 When their chromosomal positions were located, the arg mutations were found to
cluster in three areas.
  Beadle and Tatum studied each enzyme in the arginine biosynthetic pathway.
 They found that a mutation in gene cluster I made defect in enzyme E that controlled the
conversion of Gluarnate into Ornithine.
 A mutation in gene cluster 2 made defect in enzyme F that controlled the conversion of
omithine to citrulline.
 Similarly a mutation in gene cluster 3 made defect in enzyme F and enzyme H which
controlled the conversion of citruline to argininosuccinate and arggininosuccinate into
arginine respectively.
 Thus, each mutant had a defect in a single enzyme, caused by a mutation at a single site
on one chromosome.

One-gene one-polypeptide hypothesis

Each gene produces its effect by controlling the synthesis of a particular enzyme.
This is called one-gene one-enzyme hypothesis.

Since each enzyme is formed of polypeptides so this hypothesis may be known as one-gene one-
polypeptide hypothesis.
 HOW DNA-ENCODES PROTEIN STRUCTURE
DNA contains information for the maintenance of amino acid sequence in proteins.

Gene:
The sequence of nucleotides that determines the amino acid sequence in a protein is called a
gene.

CENTRIBUTION OF SANGER:

 In 1953, F.Sanger, described the complete sequence of amino acids of insulin.

 It indicated that proteins consist of fixed sequences of amino acids.

WORK OF VERNON INGRAM:

Ingram in 1956, discovered the molecular basis of sickle cell anemia.

Causes:
It occurs due to the change of glutamic acid into valine at position 6 from N terminal end of
haernoglobin β chain.

Effect:
 It reduces ability of hemoglobin to carry oxygen.
 Central dogma
The mechanism of reading and expressing of genes is referred as central dogma.

It consists of two main steps:

i. Transcription: Synthesis of mRNA from DNA.
ii. Translation: Synthesis of protein at ribosomes using information from mRNA.

Three types of RNA:

Feature mRNA tRNA rRNA

Takes message from Transfers amino acids Formation of
Function
DNA to ribosomes to ribosomes ribosomes
Single strand of Length of 75-90 Double helix with
Length
variable length nucleotides constant length
Percentage 3-4% 10-20% 80%
It is component of
- It contains codon. It contains anti codon.
ribosomes.
 Transcription
Formation of RNA from DNA with the help of RNA polymerase is called transcription.

 It is first step of gene expression.

Transcription Bubble:
Before transcription, the DNA double helix must unwind near the gene that is getting
transcribed with help of RNA polymerase. The region of opened-up DNA is called a
transcription bubble.
Process of transcription:
It involves three steps.
i) Initiation ii) Elongation iii) Termination

Initiation:
As RNA is formed from double stranded DNA (gene), only one of its strand is transcribed.
So, there are two types of DNA strands.
i) Template or antisense strand ii) Coding or sense strand

Template or antisense strand Coding or sense strand

The strand of DNA that is transcribed is The strand other than template strand is called
called template or antisense strand. coding strand.
It runs from 5’ to 3’. It runs from 3’ to 5’.
It contains anticodons. It contains codons.
Its sequence of nucleotides is opposite to Its sequence of nucleotides is same with
newly formed mRNA . respect to newly formed strand of mRNA.

Prokaryotic promoter binding site Eukaryotic promoter binding site

TTGACA -35 sequence TTGACA -75 sequence
TATAAT -10 sequence TATAAT -25 sequence
Types of RNA polymerase:
Prokaryotic RNA polymerase Eukaryotic RNA polymerase
Only one type of RNA polymerase Three types of RNA polymerases synthesize
synthesizes mRNA, tRNA & rRNA.
 RNA polymerase I makes rRNA.
 mRNA  RNA polymerase II makes mRNA.
 tRNA  RNA polymerase III makes tRNA.
 rRNA.
Initiation:
Transcription starts when RNA polymerase binds with specific sequence of nucleotides present
on template strand in start of gene. This specific sequence of nucleotides is called promoter.
Structure RNA polymerase:
RNA polymerase has two components:
i) Sigma factor ii) Core enzyme
RNA polymerase – Sigma factor = Core enzyme
 Sigma factor is responsible for correct initiation of transcription.
 Once transcription is started, sigma factor is released. The remaining molecule is called
core enzyme.

Elongation:
RNA polymerase keeps on adding ribo-nucleotides against template strand of DNA because
only thiamine ‘T’ is replaced with uracil ‘U’.

This process is continued before termination sequenced reached.

Termination:
 The stop sequence at the end of the gene stops the synthesis of mRNA.
 This stop sequence causes the RNA polymerase to detach from template strand.
  The simplest stop signal is a series of GC base pairs followed by a series of AT base
pairs.
 The RNA formed in this region forms a GC hairpin followed by four or more U
ribonucleotides.
Post transcriptional modications:
Prokaryotic mRNA Eukaryotic mRNA
In prokaryotes (bacteria) the newly In eukaryotes mRNA has to travel long
synthesized mRNA is directly released into the distance from nucleus to ribosomes.
cytoplasm.  Ribosomes are present in the
 In cytoplasm, mRNA guides the cytoplasm.
synthesis of polypeptide chain.

Modification in Eukaryotes:
 Newly synthesized mRNA is primary mRNA containing intron as well as axon.
 As intron don’t code for any amino acid, so they are spliced.
 After splicing introns, exons are later joined by RNA ligase.

Addition of Cap and Tail:

(a) Addition of a Tail:
 The tail is in the form of poly A tail linked to 3’ end of the mRNA.
 It is 100 —2000 nucleotides long.
(b) Addition of Cap:
 Cap is in form of 7 methyl GTP linked to 5’ to 5’ with first nucleotide.
 It is 300 —500 adenine nucleotides long.
Significance of modification:
These cap and tail save the mRNA from a variety of nucleases and phosphatases
in nucleoplasm and cytoplasm.

Genetic code:
Sequence of three nucleotides (arranged in triplets) that specify a particular amino acid is called
codon.
Degeneracy of codon:
Some of the amino acids are coded by more than one codon. This is called degeneracy of codon.
e.g. Isoleucene
Types of genetic code:
Case # 1
In form of single codon, there will be four types of genetic codes for 20 amino acids.
A: G = C: U
Case # 2
In form of double codon, there will be 16 types of genetic codes for 20 amino acids.
 A G C U
A AA AG AC AU
G AG GG CG UG
C AC GC CC UC
U AU GU CU UU
Case # 3
In form of triple codon, there will be 64 types of genetic codes for 20 amino acids.
Types ofcodon:
There are two types of codon.
i) Stopping or sense codons
ii) Stopping or anti sense codons codons

Stopping or sense codons Stopping or anti sense codons codons

They are present in start of mRNA. They are present at end of mRNA.
They code for methinine amino acid. They don’t code for any amino acid.
e.g. Methionine = AUG e.g. UAA, UGA, UAG

Features Genetic code is universal Genetic code is not universal

CODON NORMAL MITOCHONDRIAL DNA
UGA Stop Codon Reads tryptophan
AUA Isoleucine Methionine
AGA Reads Arginine Termination of protein synthesis
AGG Reads Arginine Termination of protein synthesis

Tranlation
This is a process in which protein is synthesized by
the ribosome according to the message present in mRNA.
Requirements:
(i) Amino acyl-tRNA
(ii) Ribosomes
(iii) Enzymes and other factors.
(iv) mRNA
(v) Factors
 Initiation factor
 Elongation factor
 Termination factor
(1) Amino acyl-tRNA:
The attachment of amino acid to 3’ end of tRNA with the help of amino acyl-RNA
synthetase (also known as activating enzyme) is called is called “Amino acyl-tRNA”.
 There are 45 different kinds of tRNAs each carrying specific amino acid.
(ii) Ribosomes:
 These are the factories for protein synthesis.
  Each ribosome has three sites named as
i) A site ii) P site iii) E site.

A site (amino acyl site) P site (Peptidyl site) E site (Exit site)
It is the site of attachment It is the site where peptide It is the site of removal of vacant
of new amino acyl-tRNA linkage will form between the (RNA molecules, which have given
molecules. two adjacent amino acids. their respective amino acids to the
growing polypeptide chain).
At this point second amino At this point first amino acyl- At this point, empty tRNA releases
acyl-tRNA molecule binds. tRNA molecule binds. out.

STEPS OF TRANSLATION:
The process of translation, in prokaryotes can be divided into three stages:
i) Initiation complex & Initiation ii) Elongation iii)
Termination
Initiation complex:
(i) Formation of initiation complex:
In prokaryotes, polypeptide synthesis begins with the formation of initiation complex.
Attachment f-Met tRNA to Ribosome:
First a tRNA molecule carrying a chemically modified methionine (called N
formyl methionine) binds to the small ribosomal subunit.
 This attachment is guided by a protein called initiation factor (IF).
IF + f-Met tRNA = Initiation complex
Attachment of initiation complex to mRNA:
This initiation complex, guided by another initiation factor, binds to starting codon “AUG” on
the mRNA at P site.
2. INITIATION:
Attachment of large subunit:
 Large ribosomal subunit binds to smaller one.
  Initiation factors are released and neutralized.

2. ELONGATION:
(i) Binding of new amino acyl tRNA:
When a new amino acyl tRNA with the appropriate
anticodon appears, proteins called elongation factors (EF)
assist in binding it to the exposed mRNA codon at the A site.

(ii) Formation of peptide bond:

 N-formyl methionine detaches from its tRNA (at P-site)
and attaches to the second amino acid present on A site.
 This reaction is catalyzed by large ribosomal subunit.

(iii) Translocation and liberation of free RNA:

 The ribosome now moves (translocates) three more nucleotides (a codon) along the
mRNA molecule in the 5’ —3’ direction.
 This movement is guided by other elongation factors.
 This movement translocates the initial tRNA to the E site and releases it from the
ribosome.
 Such movement keeps on translocating until the stop codon is reached.

4. TERMINATION:
Elongation continues until the stop codon is reached (for example UAA. UGA or UAG).
 These stop codons are recognized by recognized by release factors (RF).
 The release factors are the proteins that release the newly made polypeptide from the
ribosomes.
 Mutation
 A gene mutation is a permanent change in the DNA sequence that makes up a new allele
in the population.
 Mutations range in size from:
 Change in a single DNA nucleotide.
 Change in large segment of a chromosome.
 Change in whole chromosome.
 Change in the number of chromosomes.
If the DNA in all of the cells of an adult human were lined up end to end, it would stretch nearly
100 billion kilometer — 60 times the distance from Earth to Jupiter.
Mutagens Mutant
The agents that causes mutations are called The organism or cell in which mutation is
mutagens occurred is called mutant.
e.g. chemical compounds or radiation (such as
UV or X-rays)
 causes irreversible and heritable
changes (mutations) in DNA.
  The mutations in somatic cells do not pass on to offspring.
Such mutations have little evolutionary consequence.
 Mutation in germ line cell is passed to next generations.
Such mutations providing the raw material for evolutionary change through natural
selection.
Types of Mutations:
Generally, there are two types of mutations
i) Chromosomal aberration ii) Point mutation
Chromosomal Mutations:
“Changes in structure or number of chromosomes are referred as chromosomal mutations
or chromosomal aberrations.”
i) Change in no. of chromosomes:
The changes appear as a result of unequal division of homologous chromosomes
during meiosis, termed as non-disjunction of chromosomes.
Example:
 Down’s syndrome.
 klinefelter’s syndrome.
 Turner’s syndrome.
ii) Change in structure of chromosomes:
These are the changes within the segment of chromosomes and are of following types.
Deletion:
The loss of a segment of a chromosomes or of a part of the gene.
Duplication:
When a part of chromosome is repeated or present more than once, it is called duplication.
Inversion:
Where a part of chromosome becomes inverted.
Transposition or Translocation:
Transfer of a segment of chromosome to a non-homologous chromosome is called translocation.

Gene/Point Mutations:
 A gene or point mutation arises as a result of a chemical change in an individual gene.
Causes:
a) Spontaneous pairing errors that occur during DNA replication.
b) Some other mutations result from damage to the DNA.
Effects:
A change in the sequence of nucleotides results in change in order of amino acids in protein.
Examples:
Sickle cell anemia and phenylketonuria are very well known examples of point mutations.
 Sickle Cell Anemia Phenylketonuria
Definition: Definition:
Alteration in the tertiary structure of the A rare condition in which a baby is born
haemoglobin molecule reducing its ability to without the ability to properly break down an
carry oxygen is called sickle cell anemia. amino acid called phenylalanine is called
phenylketonuria.
Causes: Causes:
It occurs due to the change of glutamic acid It occurs due to absence of phenylalanine
into valine at position 6 from N terminal end hydroxylase.
haernoglobin β chain.
Effects: Effects:
This reduces its ability to carry oxygen. Consequently, Phenylalanine accumulates in
brain cells leads to mental retardation.