Nucleic Acid

Nucleic Acid
NUCLEIC ACID
• Initially, nucleic acid was known by the term nuclein by Friedrich
Miescher, a 25 year old swiss chemist, isolated nuclei from pus cells
(white blood corpuscles) and found that they contain a unknown
phoshate rich substance, which he named nuclein.
• The nucleic acids are the heredity determinants of living organisms.
• A nucleic acid is a chain of nucleotides which stores genetic information
in biological systems.
• There are two types of nucleic acid: deoxyribonucleic acid (DNA) and
ribonucleic acid (RNA).
 Deoxyribonucleic acid (DNA) is a self-replicating
material which is present in nearly all living
organisms as the main constituent of chromosomes.
 found in nucleus, mitochondria and chloroplasts
 carries the genetic information.
•Components:
a) Nucleic acid/ nitrogenous bases
(adenine, thymine, cytosine and guanine).
b) a five carbon sugar called deoxyribose,
c) a phosphate molecule
The Secondary structure of DNA
Double Helical model of DNA
 In 1953, Watson and crick proposed universally accepted famous
double helical model for the structure of DNA.
 The features of double helix model are:
 DNA molecule is composed of two complementary polynucleotide
chain or strand.
 Each nucleotide strand is made up of deoxyribonucleotides jointed by
phosphodiester linkage.
 Two strands of DNA are oriented anti-parallel to each other i.e 5’ end
of one strand is located with 3’ end of other strand towards the same
end of DNA.
 The sugar phosphate linkage forms the backbone of the double helix.
The nitrogen bases form the inside portion i.e. core of double helix.
 Adenine pair with Thymine of other chain, while G with C.
 Follows Chargaff’s rule (A=T & G = C also
A+G=T+C).
 Pairing of bases between two strand occurs by
hydrogen bonding. A=T form double bond & G Ξ
C form three hydrogen bond between two strand.
 Two strands are twisted around each other on
common axis forming a right handed helix.
 The genetic information resides on one of the
two strands known as template strand or sense
strand. The opposite strand is called antisense
strand.
 Diameter- 20A°, One complete turn-34A°,
distance between two bases-3.4A°
Forms of DNA
B-Form of DNA (B-DNA):
• Proposed by Watson and Crick. It is present at a very high relative humidity (92%) and
low concentration of ions. It has antiparallel double helix, rotating clockwise (right
hand) and made up of sugar- phosphate back bone combined with base pairs or
purine-pyrimidine. The base pairs are perpendicular to longitudinal axis of the helix.
The base pairs tilt to helix by 6.3°. The B-form of DNA is metabolically stable.
A-Form of DNA (A-DNA):

• The A-form of DNA is found at 75% relative humidity in the presence of Na+, K+ or Cs+
ions. Contains eleven base pairs as compared to ten base pairs of B-DNA which tilt from
the axis of helix by 20.2°.
C-Form DNA (C-DNA):

• The C-form of DNA is found at 66% relative humidity in the presence of lithium (Lit+)
ions. As compared to A-and B-DNA, in C-DNA the number of base pairs per turn is less
i.e. 28/3. The base pairs show tilt by 7.8°.
D-Form of DNA (D-DNA):
• The D-form of DNA is found rare. Total number of base pairs per turn of
helix is eight. Therefore, it shows eight-fold symmetry.
Z-Form of DNA (Z-DNA):

• Left handed double helix model with zig-zag sugar-phosphate back
bone running in antiparallel direction. Therefore, this DNA has been
termed as Z-DNA. The Z-DNA has been found in a large number of living
organisms including mammals, protozoans and several plant species.
HISTORY
• Fredrick Griffith in 1928 performed experiment in s. pneumoniae earlier
called Diplococcus pneumoniae
• Two strain of streptococcus pneumoniae used in his experiment.
Virulent strain Avirulent strain

(Smooth colony) (Rough colony)
enclosed in lipopolysaccharide
lacked a Lipopolysaccharide (LPS)
capsule
capsule
kill mice by causing disease Mice survive & Do not cause

Pneumonia pneumonia
Injecting strains in different ways in the mice and obtain following results:
i ii iii iV
Griffith’s experiment conclusion
• Experiment i, ii and iii gave expected result but experiment iv result
was unexpected.
• When heat killed smooth strain and live rough strain are injected
together it causes pneumonia and mice dead. After analysis of dead
mice live smooth strain obtained in body.
• Griffith concluded there must be some material in heat killed
smooth strain that changed rough strain to smooth strain.
• He called this phenomenon transformation.
• His experiment cannot provide the material that change rough strain
to smooth strain.
Avery, MacLeod, & McCarty experiment
• Oswald Avery, Colin McLeod, and Maclyn McCarty ran an

experiment based on Griffith's work in 1944 and made a major
breakthrough proving that DNA was genetic material and its ability
to transform.
• Experiment was conducted in vitro, in lab & are summarized as:
 They isolated extracts from the heat-killed S-strain and manipulated
them to get rid of proteins, RNA, and DNA.
 The R-strain was then added to the isolated extracts.
 Not all extracts transformed into a strain that caused death in mice.
Avery, MacLeod, & McCarty experiment
Experiment was conducted in vitro, in lab & are summarized as:
1. Rough/Avirulent Strain Culture Rough colony
2. Smooth strain Culture Smooth colony
3. Heat killed smooth strain culture No colony
4. DNA isolated smooth strain culture No colony
5. Rough strain (live) Culture Smooth colony
+ Heat killed smooth strain
6. Rough strain (live) Culture Smooth colony
+ DNA of smooth strain
 From above experiment , it was observed that DNA is the transforming
factor. But isolated DNA contain some trace of RNA and protein. So
further experiment was conducted.
From above experiment it clear that:
 Protein was digested by protease and RNA was digested by Rnase , still
smooth colony was formed
 When DNA was digested by DNase, No colony was formed.
 This experiment prove that DNA is the transforming element.
Hershey And Chase Experiment
• Alfred Hershey and Martha Chase, 1952 investigated bacteriophages:
(viruses that infect bacteria)
- the bacteriophage was composed of only DNA and protein
- they wanted to determine which of these molecules is the genetic
material that enters into the bacteria.
• They studied the life cycle of T2 phage of E. Coli
HERSHEY & CHASE (1952) EXPERIMENT:
• In culture I : Bacteriophage was grown in medium containing
Radioactive Phosphorus (32P) to make DNA Radioactive
• In culture II : Bacteriophage was grown in medium containing
radioactive Sulphur (35S) to make proteins Radioactive
• Both kinds of Bacteriophage particles were allowed to infect Bacteria
• The infected bacteria were observed for radioactivity
• Radioactive phosphorus was found with bacterial cells but Radioactive
Sulphur was not traced in bacterial cells ( Only in Ghosts)
• Bacteriophage progeny carried only radioactive phosphorus and not
radioactive sulphur.
Hershey And Chase Experiment
Hershey And Chase conclude:
 As ghosts or coat of Bacteriophage were not labelled with 32P and
only with 35S.
 The results of experiment clearly indicate that only DNA and not the
proteins enter the bacterial cell. Protein coat is left outside.
 The DNA entering the host cell carries all the genetic information for
synthesis of new phage particle.
 This certainly proves that DNA is the genetic material in
Bacteriophage and not proteins

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Nucleic Acid

A-Form of DNA (A-DNA):

C-Form DNA (C-DNA):

Z-Form of DNA (Z-DNA):

Virulent strain Avirulent strain

kill mice by causing disease Mice survive & Do not cause

• Oswald Avery, Colin McLeod, and Maclyn McCarty ran an

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