ZOOLOGY UNIT 7: CHAPTER 5.
MOLECULAR BASIS OF INHERITANCE:
IV. REPLICATION OF DNA:
Definition: Replication of DNA may be defined as a process by which a DNA molecule forms exact copies
of itself.
DNA replication takes place during S-phase (synthesis phase) of the interphase stage of the cell cycle
(Figure 1)
FIGURE 1. DNA REPLICATION IN S PHASE (SYNTHESIS PHASE) OF THE INTERPHASE.
MECHANISM OF DNA REPLICATION: (Refer to figure 2)
The entire process of DNA replication involves the following steps:
1. Activation of deoxyribonucleotides: The four nucleotides of DNA such as dAMP (deoxyy
adenosine monophosphate), dGMP (deoxyy guanosine monophosphate),dCMP (deoxyy cytidine
monophosphate) and dTMP ((deoxy thymidine monophosphate) are activated by ATP to form
ATP,GTP,CTP and TTP.
ATP
dAMP ATP
ATP
dGMP GTP
ATP
dCMP CTP
ATP
dTMP TTP
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2. Recognition of initiation point: Replication of DNA begins at a specific point called initiation
point or origin. Specific initiator proteins recognize or identify the initiation point on the DNA
molecule.
3. Unwinding of DNA molecule: The double stranded DNA helix uncoils to form single stranded
DNA by breakage of weak hydrogen bonds. The enzyme helicase helps in breaking the hydrogen
bonds between two strands of DNA and the enzyme topoisomerase cut one strand of DNA helping
the separation of the two strands of DNA. Thus, unwinding of DNA gives a Y-shaped structure to
the two strands of DNA duplex which is known as Replication fork.
4. Formation of RNA primer: Primer is a short segment of RNA which is formed on the DNA
template before replication begins. The enzyme primase is involved in the formation of RNA
primer. (The RNA primers are complementary to the two strands of DNA and formed of 50- 100
nucleotides).
5. Synthesis of new DNA strands on RNA primers: One new strand of DNA is formed in a
continuous stretch in 5/→3/ direction from the 3/→5/ template due to the addition of
deoxyribonucleotides at the 3/ end of primer RNA. This process occurs in presence of the enzyme
DNA polymerase and ATP the strand thus formed is called Leading strand.
After the synthesis of leading strand, the same enzyme (DNA polymerase) forms short
segments of DNA on the other parent strand again in 5/→3/ direction, each starting from an RNA
primer. These short segments of DNA are called Okazaki fragments.
6. Removal of RNA primers: After the formation of okazaki fragments, RNA primers are removed
from the 5/ ends one by one by the exonuclease activity of the DNA polymerase, leaving gaps in the
newly synthesized deoxyribonucleotide strand.
7. Joining of Okazaki fragments: The gaps left between the okazaki fragments are filled with
complementary deoxyribonucleotide residues by DNA polymerase-I. Finally, the adjacent 5/ and 3/
ends are joined by the enzyme DNA LIGASE. The strand formed as a result of joining of these
fragments is called Lagging strand.
8. Proof reading and DNA repairing: The base pairing in DNA is highly specific. During DNA
replication, if wrong bases enter into the helix of DNA, the enzyme DNA polymerase removes
them. This process of removal of wrong bases so as to ensure accurate replication is called PROOF
READING.
The wrong bases entered into the helix of DNA by mutation can be identified and corrected
by certain enzymes called repair enzymes such as NUCLEASES. These enzymes cut off the defective
segments of DNA and at the same time, introduce and join the normal correct segment by the
enzyme [Link] entire process is called DNA REPAIRING
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DNA MOLECULE
HYDROGEN BOND
BETWEEN TWO DNA
STRANDS
REPLICATION
BREAKS HYDROGEN BONDS
FORK
PARENTAL STRAND
DAUGHTER
STRANDS
PARENTAL STRAND
FIGURE 2. MECHANISM OF DNA REPLICATION.
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WATSON AND CRICK'S SEMI-CONSERVATIVE MODE OF DNA REPLICATION:
Watson and Crick suggested a simple mechanism of DNA replication on the basis of its double
helical structure known as semi-conservative method of DNA replication.
Explanation:
As shown in the figure 3, the new DNA strands (daughter strands) formed as a result of
replication are half old and half new. In other words, half of the original molecule is conserved and
its complementary half is newly synthesized according to the sequence of bases contained in the
conserved half. Therefore, this method of DNA replication is known as semi-conservative
method of DNA replication.
Half of the original
(parental) strands
are conserved
DAUGHTER STRANDS Newly synthesized
FIGURE 3. WATSON AND CRICK'S SEMI-CONSERVATIVE METHOD OF DNA REPLICATION.
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REVISION QUESTIONS:
(A) Very short answer type:
i. What do you mean by replication of DNA? 1×6=6
ii. When DNA replicates?
iii. Who proved experimentally that DNA replication is semi conservative in nature?
iv. Name the enzyme which helps in unwinding of DNA molecule?
v. Which enzyme is responsible for cutting one strand of DNA thereby helping the separation of two
strands of DNA duplex?
vi. What do you mean by “semi conservative” mode of DNA replication?
(B) Fill in the blanks: 1×4=4
(i) Replication of DNA begins at a specific point called ----------- point or------------.
(ii) The enzyme involved in the synthesis of RNA primer is-----------.
(iii) The process of formation of new DNA chains or RNA primers occurs in the presence of the enzyme--
---------.
(iv) The short DNA segments formed in the 5’- 3’direction starting from RNA-primers are known as-----
--------.
(C) Short answer type:
What is the necessity for self duplication of DNA & chromosomes? 2
(D) Long answer type:
Give an account of the mechanism of DNA replication. 4/5
(Add diagrams to explain the different steps)