DNA Replication

• An essential feature of DNA is that it must be able to replicate itself

accurately , so that when a cell divides, the genetic code it carries can be
passed to the daughter cells.

• DNA replication copies precisely so that the new molecules are produced with
exactly the same sequence of bases as the original strands.

• DNA replication takes place in the nucleus during interphase of the cell cycle
when DNA is not tightly coiled.
Steps of replication
1. The first step in the process is the ‘unzipping’ of the two strands. Helicase
moves along the double helix, unwinding the two strands, which separate from
one another as the relatively weak hydrogen bonds between the bases are
broken

2. The unpaired nucleotides are exposed and each single strand now acts as a
template for the formation of a new complementary strand. Free nucleotides
move into place: C pairs with G and A pairs with T.

3. The free nucleotide bases form complementary pairs with the bases on the
single DNA strands. DNA polymerase is the enzyme involved in linking the new
nucleotides into place. Finally, the two new DNA molecules are rewound, each
one forming a new double helix.
ENZYMES THAT ARE INVOLVED IN DNA REPLICATION IN EUKARYOTES
• Helicase: Unwinds and separate the two strands of DNA by breaking the hydrogen
bonds.
• Primase: adds the short portion of RNA called primers at the start of the leading and
sections of the lagging strand where DNA polymerases start the process of
replication.
• DNA Polymerase III: adds nucleotides one by one on the leading strand and in
chanks called Okazaki fragments on the lagging strand.
• DNA Polymerase I: Removes the RNA primers after a new DNA strands have been
synthesised.
• Exonucleases: these enzymes acts as a proof readers during DNA polymerisation
(synthesis of the new strand) during replication by removing wrong nucleotides and
are involved in repairing the DNA.
• Ligase: Plays an important role of joining or silling the breaks or gaps in the
phosphodiester backbone of DNA that occur during replication and recombination,
and as a consequence of DNA damage and its repair.
Synthesis of a new DNA as the original
replicates
The figure below shows replication
• DNA polymerase III move along the template strand in the same direction, adding one
nucleotide at a time
• Free nucleotides with each of the four possible bases are available in the area where DNA is
being replicated
• Every time a nucleotide is added to the new strand only one of the four types of nucleotide has
the base that can pair with the base at the position reached on the template strand.
• DNA polymerase brings nucleotides into the position where hydrogen bonds could be formed
but unless this happens and a complementary base pair is formed, the nucleotide break away
again
• Nucleotide is finally has the correct base and has been brought into position and hydrogen
bonds have been formed between the two bases, DNA polymerase links it to the end of the new
strand
• This is done with Covalent bonds between the phosphate group of the free nucleotide and the
sugar of the nucleotide at the existing end of the new strand
• Pentose sugar is 3 terminal and the phosphate is the 5 terminal, DNA polymerase adds on the 5
terminal of the free nucleotide to the 3 terminal of the existing
• DNA polymerase continues to move along the template strand creating new strands with a base
sequence complementary to the template strand – it does this with a very high degree of fidelity
(very few mistakes made)
DNA replication being a semi-conservative process
DNA replication is a semi-conservative process, because when a new double-stranded DNA
molecule is formed:
• One strand will be from the original template molecule.
• One strand will be newly synthesised
• When a cell is preparing to divide, the two strands of the double helix separate. The new
strands are used as a guide or template for the creation of a new strand.
• New strands are formed by adding nucleotides, one by one, and linking them together
• The results of this process is 2 DNA molecules, both made up of the original strand and a
newly synthesized strand.
• Therefore DNA replication is referred to as being semi-conservative. DNA replication is said to
be semi-conservative because no DNA molecule is ever completely new.
• Every double helix contains one ‘original’ and one ‘new’ strand.
The base sequence on the template strand determines the base sequence on the new strand,
only a nucleotide carrying a base that is complementary to the next base on the template
strand can successfully be added to the new strand.
• Since complementary bases form hydrogen bonds with each, stabilizing the
structure, if a nucleotide with the wrong base started to be inserted, the hydrogen
bond would not happen and the nucleotide would not be added to the chain
Rule – one base always pairs with another is called complementary base pairing.
This makes sure that the two DNA molecules that are created by DNA replication are
identical in their base sequences to the parent molecule that was replicated.

Class exercise
• Describe the procedure of the Meselson and Stahl experiment.
• Explain how the Meselson and Stahl experiment demonstrated semi-
conservative DNA replication.