DNA as a Genetic Material

Transforming Principle:

Frederick Griffith in 1928, carried out a series of experiments with Streptococcus pneumoniae (a bacterium that
cause pneumonia). He observed that when these bacteria (Streptococcus pneumonia) are grown on a culture plate,
some of them produce smooth, shiny colonies (S-type), whereas, the others produce rough colonies (R-type). This
difference in character (smooth/rough) is due to a mucous (polysaccharide) coat present in the S-strain bacteria,
which is not present in the R-strain.

In his experiments, he first infected two separate groups of mice. The mice that were infected with the S-strain die
from pneumonia.

‘S’ strains are the virulent strains causing pneumonia.

The mice that were infected with the R-strain do not develop pneumonia and they live.

S-strain (virulent strain) → Inject into mice → Mice die

R-strain (non-virulent strain) → Inject into mice → Mice live

In the next set of experiments, Griffith killed the bacteria by heating them. The mice that were injected heat-killed S-
strain bacteria did not die and lived, whereas the mice that were injected a mixture of heat-killed S-strain and live R-
strain bacteria, died due to unexpected symptoms of pneumonia.

S-strain (heat killed) → Inject into mice → Mice live

S-strain (heat killed) + R-strain (live) → Inject into mice → Mice die

Griffith concluded that the live R-strain bacteria, were transformed by the heat-killed S-strain bacteria.

He proved that there was some ‘transforming principle’ that was transferred from the heat-killed S-strain, which
helped the R-strain bacteria to synthesise a smooth polysaccharide coat and thus, become virulent. That was due to
the transfer of the genetic material.

However, he was not able to define the biochemical nature of genetic material from his experiments.

Biochemical Characterisation of Transforming Principle:

Oswald Avery, Colin MacLeod and Maclyn McCarty (1933-44) worked to determine the biochemical nature of
‘transforming principle’ in Griffith’s experiment in an in vitro system.

From the heat-killed S-cells, they purified biochemicals (proteins, DNA, RNA, etc.) to observe, that which
biochemicals could transform live R-cells into S-cells.

Therefore, they discovered that DNA alone from heat-killed S-type bacteria caused the transformation of non-
virulent R-type bacteria into S-type virulent bacteria.
Protein-digesting enzymes (proteases) and RNA-digesting enzymes (RNases) did not cause this transformation. This
proved that the ‘transforming substance’ was neither the protein no RNA.

DNA-digesting enzyme (DNase) caused inhibition of transformation, which suggests that the DNA caused the
transformation. Thus, these scientists came to the conclusion that DNA is the hereditary material.

STRUCTURE OF NUCLEIC ACIDS (DNA AND RNA)

All living organisms possess, nucleic acids. Nucleic acids are macromolecules of utmost biological importance.
Nucleic acids possess all the information needed for an organism’s cell structure, function, development and
reproduction.

In 1889 Altmann coined the term nucleic acid. Later existence of two types of nucleic acid, deoxyribonucleic acid
(DNA) and ribonucleic acid (RNA) was established.

Each nucleotide is composed of three distinct molecules : one molecule each of sugar, phosphoric acid and a
nitrogenous base.

Sugar:

All nucleotides contain a 5-carbon sugar (pentose); the pentose ribose is found in RNA while deoxyribose is found in
DNA. In deoxyribose molecules, one oxygen atom (O) is missing from 2′ position (Fig. 3.1). The nucleic acids (NA) are
named after the sugar present in them, for example,

Phosphoric Acid:

Phosphoric acid (H3PO4) is attached to each sugar at the 3′ and 5′ C positions to give rise to the sugar-phosphate
backbone. Free nucleotides in the cell have 3 phosphate residues, generally attached to the 5′ C of the pentose.
During the phosphodiester bond formation, two phosphate groups are removed from one of the two participating
nucleotides.
Nitrogenous Bases:

The bases in nucleic acids are heterocyclic compounds containing nitrogen and carbon in their rings. The nitrogenous
bases are of two types: pyrimidines and purines.

Pyrimidines:

Pyrimidine ring is similar to the benzene ring, except it contains nitrogen in place of carbon at positions 1 and 3 (Fig.
3.3). They also contain a keto oxygen (=0) at the position 2. There are three common pyrimidines cytosine (C),
thymine (T) and uracil (U).

Thymine contains two ketooxygens at positions 2 and 6 and a methyl group (-CH3) at position 5. Cytosine contains
one keto oxygen at position 2 and an amino group (-NH2) at position 6. These two pyrimidines are found in DNA,
while another pyrimidine uracil occurs in RNA in the place of thymine. Uracil differs from thymine only in not having
a methyl group at the position 5. Pyrimidines are associated with 1′ C of the sugar by the position 3.

Purines:

Purines have two carbon-nitrogen rings. One of the rings is 6 membered (like pyrimidine), while the other is 5
membered; the two rings share their 4 and 5 C (Fig. 3.3). Both RNA and DNA contain the same two types of purines,
viz., adenine (A) and guanine (G).

Adenine contains an amino group (-NH2) at position 6, while in guanine this position is occupied by a keto oxygen
(=0). In addition, guanine has an amino group at position 2. Both the purines contain nitrogen at positions 1, 3, 7 and
9. Purines associate with 1′ C of pentose sugar at their position 9 N.

Nucleosides:

The combination of a base and a pentose is termed as nucleoside. The 1′ C of pentose attaches to the 3-position of a
pyrimidine or at the 9-position of a purine. Nucleosides derived from ribose are called ribosides, while having de-
oxy-riboseare known as de-oxy-riboside; the various nucleosides are as follows:
Nucleotides:

When a phosphoric acid molecule is attached to the pentose residue of a nucleoside, it is called a nucleotide.
Phosphoric acid may attach at either 5’C or 3′ of the pentose; accordingly the nucleotides are called either 5′ P3′ OH
nucleotides or 3′ P5′ OH nucleotides.

However, only the 5′ P3′ OH nucleotides occur naturally. There are four different ribonucleotides (ribotides) as well
deoxyribonucleotides (de-oxy-ribotides). The free nucleotides present in the cells are found in triphosphate form,
e.g., ATP, GTP, TP, UTP (ribonucleotide triphosphates) and dATP, dGTP, dCTP and dTTP (deoxyribonucleotide i
phosphates).