Genetic Material (DNA and RNA)

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Gene:
It is a hereditary unit which is transmitted from one generation to
another generation.
Properties and function of gene:
1.Genes are present in a lengthwise fashion on the chromosomes
which has at least two alleles.
2.It occupies a specific locus on a chromosomes.
3.Gene determine the phenotype and genetic constitution of organism.
4.Genes express themselves through the production of chemical
substances.
Griffith’s experiment:
In 1928 Frederick Griffith was interested in developing a vaccines
against the Pneumococcus which causes pneumonia. One strain has a
capsules and can causes pneumonia which is smooth. The harmless
strain is rough. Griffith experiment obtained the following results.
Experiment 1: Living rough Pneumococcus bacteria was injected. It
did not causes pneumonia. Mice remained healthy. Rough
Pneumococcus bacteria are not injective.
Experiment 2: Living smooth Pneumococcus bacteria was injected. It
caused pneumonia. Mice died. Smooth Pneumococcus bacteria are
injective.
Experiment 3: Heat killed smooth Pneumococcus bacteria was
injected. It did not causes pneumonia. Mice remained healthy. Smooth
Pneumococcus bacteria that are killed by heat are not injective.
Experiment 4: Living rough and heat killed smooth Pneumococcus
bacteria were injected together. Mice suffer from pneumonia and died.
On the basis of these results, he concluded that ‘ something must be
passing from the heat killed capsulated forms which caused them to
develop capsules and become virulent.
 Fig. Griffith’s experiment on transformation of bacteria.

Further in 1944, Oswald Avery, Colin Macleod and Maclyn McCarty

elaborated Griffith’s experiment in order to identify transferring
substances which obtained the following results;
1.Living non-capsulated Pneumococcus + capsule or heat killed
capsulated bacteria → Mice remain alive.
2.Living non-capsulated Pneumococcus + cell wall of heat killed
capsulated bacteria → Mice remain alive.
3.Living non-capsulated Pneumococcus + DNA of heat killed
capsulated bacteria → Mice get infected by pneumonia and died.
4.Living non-capsulated Pneumococcus + DNA of heat killed
capsulated bacteria + enzyme deoxyribonuclease → Mice remain alive.
From the experiment, they strongly recommended that DNA is the
genetic material and is responsible for the transformation in bacteria.

Deoxyribonucleic Acid (DNA):

DNA is mainly found in the nucleus of eukaryotic cell, but also found in
mitochondria and chloroplast in little amount. It is formed by the end
to end polymerization of a large number of repeated units called
nucleotides. Those nucleotides are formed by i) deoxy-ribose sugar ii)
Phosphate and iii) nitrogenous base.
i) Deoxy-ribose sugar: It is a pentose sugar having 5 carbon atoms. Due
to the deoxyribose sugar it is called deoxyribose nucleic acid.
ii) Phosphate: the phosphate in the DNA is present as phosphoric acid
(H3PO4).
iii) Nitrogenous Bases: the nitrogenous bases are of two types- Purine
and pyrimidine. Purine bases comprise mainly adenine (A) and guanine
(G) while pyrimidine bases comprise cytosine (C) and thymine (T).
Nucleoside: nitrogenous base + sugar
Nucleotide: nucleoside + phosphate

Watson and crick’s model of DNA

J.D. Watson and F. crick (1953) combined the physical and chemical
bata generated by early work and purposed a double helix model for
DNA molecule. This model is widely accepted. According to this
model, the DNA molecule consists of two strand which are connected
together by hydrogen bonds and helically twisted. Each step on the
one strand consist of a nucleotide of purine base which alternate with
that of pyrimidine base. Thus, a strand of a DNA molecule is a polymer
of four nucleotide i.e. A, G, T, C. the two strand join together to form
double helix. Bases of two nucleotide form hydrogen bond i.e. A
combines with T by two H-bond (A=T) and G combines with C by three
hydrogen bond. However, the sequence of bonding is such that for
every A.T.G.C. On one strand there would be T.A.C.G. on the other
strand. Therefore, the two strand are complementary to each other.
The two strands of double helix run in antiparallel direction, i.e. they
have opposite polarity.
 Fig: Structure of DNA (Watson and Crick
Model)

The hydrogen bonds between the two strands are such that maintain a
distance of 20 A0. The double helix coils in right hand direction. The
turning of double helix results in the formation of a deep and wide
groove called major groove. The major groove is the site if bonding of
specific protein. The distance between two strands forms a minor
groove. The DNA model also suggested a copying mechanism of the
genetic material. DNA replication is the fundamental and unique event
underlying growth and reproduction in all living organism ranging from
the smallest viruses to the most complex of all creatures including
man. DNA replicates by semiconservative mechanism.

DNA replication:
The process of synthesis of DNA molecule from preexisting DNA is
called DNA replication.DNA replication may occur by following
process:

1.Semi-conservative replication
2.Conservative replication
3.Dispersive replication

Semi-conservative mode of DNA replication

Enzymes involved in semi conservative DNA replication:
Helicase- helicase enzyme breaks the hydrogen bonds formed between
bases of nucleotides
Topoisomerase- topoisomerase enzyme breaks and reseals one strand
of DNA
DNA polymerase- it connects the nucleotides to form DNA polymer
nucleotides. It forms RNA primer in DNA strand which is like a knot
formed o thread. Due to its formation, newly formed DNA strands
cannot separate.
Ligase- ligase binds the fragments of DNA strand together
Repair enzyme- this enzyme cuts off wrong base of nucleotide and
rejoin correct base in newly formed DNA

Process:
DNA replication in eukaryotes may begin at several points. In
replication ,the helicase enzyme breaks the hydrogen bond between
the bases of nucleotides. due to breaking of hydrogen bonds of
nucleotides, the two strands separate. The process of separation of
DNA strands also supported by enzyme topoisomerase. This enzyme
breaks and reseals one strand of DNA molecule.

Now the bases of DNA strands become exposed and joins with the free
nucleotides present in nucleoplasm only in 5’-3’ direction. This process
or formation of polynucleotide chain takes place in the presence of
enzyme DNA polymerase. The four types of nucleotides are present in
nucleus in the form of dATP, dGTP, dTTP and dCTP. These
molecules are triphosphates and break down into monophosphates
dAMP, dTMP, dCMP and dGMP with the release of energy. These
monophosphates nucleotides make hydrogen bonds with the
complementary nucleotides of DNA strand.
According to base pairing rule the nucleotides are arranged. As DNA
strand formation occurs only in 5’-3’ direction , in on strand of DNA a
continuous strand is formed called leading strand ehereas in other
strand of DNA, fragments of strands are formed, these fragments are
called Okazaki fragments named after Japanese scientist Okazaki.
The strand with Okazaki fragments is called lagging strand.

Here, both DNA strands take part in replication process, so they are
called as templates. The segments of strands are joined by the help of
enzyme ligase. At the beginning of strand formation, RNA primer is
formed at the DNA strand and it is due to the enzyme primase.
Sometime a wrong base maybe linked with the DNA nucleotide. When
it occurs, the repair enzyme replaces the wrong base by correct
base.
In the above process, newly formed DNA have one strand of old DNA
and one of new formed polynucleotide chain. Hence the process is
called as semi conservative process of DNA replication. It means half
of parent DNA is conserved in new formed DNA.

Structure of RNA:
The RNA is usually single stranded except viruses such as TMV,
yellow mosaic virus, reovirus etc. RNA is found both in the nucleus and
in the cytoplasm. The single strand of the RNA is folded either at
certain regions or entirely to form hairpin shaped structure. The RNA
dose not possess equal purine-pyrimidine ratio, as is found in the DNA.
 Fig: Structure
of RNA.
Like DNA, the RNA is also the polymer of four nucleotides each one
contains D-ribose, phosphoric acid and a nitrogenous base. The bases
are two purines (A,G) and two pyrimidines (C,U). Thyamine is not found
in RNA. Pairing between bases occurs as A-U and G-C.
If the RNA is involved in genetic mechanism, it is called genetic RNA
as found in plant, animal, and bacterial viruses. The DNA acts as
genetic material and RNA follows the order of DNA, In such cells the
RNA dose not have genetic role. Therefore, it is called non-genetic
RNA. The non- genetic RNA is of three types:
1. m-RNA(messenger RNA): it constitutes about 5% - 10% of the total
RNA present in the cell, m-RNA carries the genetic information from
DNA for Protein synthesis.
2. r- RNA(Ribosomal RNA): It makes about 80% of the total RNA in the
cell. It is the major component of ribosomal.
3. t RNA (Transfer RNA): it is also known as soluble RNA. It forms
about 10% - 15% of the total RNA. It used to transfer amino acid
molecule to the site of protein synthesis.
A nucleotide is made up of three molecules. These are
a) Pentose sugar: the pentose sugar is ribose.
b) Nitrogenous bases: i) Purine = Adenine And Guanine ii)
Pyrimidine=Cytosine And Uracil
c) Phosphoric Acid

Fig. Structure of t-RNA

Transcription ( RNA synthesis):

Transcription is the first step of gene expression, in which a particular
segment of DNA is copied into RNA by the enzyme RNA polymerase.
The transcript RNA is complementary to DNA in base sequence.
Transcription involves the following steps;
1.A portion of DNA strand uncoils and the two strand separate at a
specific points.
2.One strand of DNA( 3’-5’) DNA act as master strand for the formation
of RNA, while other strand ( 5’-3’) does not participate in RNA
formation.
3.The template strand has a promoter or initiation site and a
terminator site.
4.RNA polymerase initially binds at the promoter site of the template
DNA strand and proceeds the formation of RNA strand.
5.RNA synthesis initiates from 5’ to 3’ direction on the master strand.
6.Free ribonucleotides slot into the place opposite the exposed bases
on the template DNA strand.
7.RNA polymerase moves alone template DNA causing elongation of
RNA.
8.The sugars and phosphate of the m-RNA nucleotides are then linked
together to form a RNA molecule.
9.RNA gets detached and leaves the nucleus through nuclear pore into
the cytoplasm.
10.The DNA strand coil up again and formed the double helix.