DNA REPLICATION

• DNA replication is the process by which the genome's DNA is

copied in cells.

• Replication increases the DNA copies in the cells

• The entire genome is copied so that each new daughter cell

ends up with its own complete genome
PROCESS/STEPS IN DNA REPLICATION
1. Recognition of initiation point
- Starts at specific point called origin of replication (OR)
- Replication fork forms at OR
- OR is a nucleotide sequence of 100 to 200 pairs of bases.
- Specific initiation proteins recognize the initiation site on DNA
- Such proteins along with DNA directed RNA polymerase initiate the
synthesis of RNA primer
- RNA primer – responsible for the formation of DNA chain.
*IN prokaryotes – only one replication fork
*In Eukaryotes – several replication forks are formed
2. Unwinding of DNA –

• DNA duplex molecule is cut open (nicked) to form a bubble or fork

• unwinding proteins get attached at the point of nick
• Separation of the strands of the DNA duplex occurs
• Unwinding occurs in the presence of DNA HELICASE an enzyme.

3. Template DNA –
• The single stranded DNA on which the new DNA is synthesized is
called template DNA.
4. RNA Primer –

• RNA polymerase synthesizes RNA primer on template DNA

• In the absence of RNA primer the DNA replication is irregular.

5. Chain Elongation –

• New DNA strand is formed due to DNA polymerase III enzyme

• This enzyme adds nucleotides in 5’ to 3’ direction
• This activity of DNA polymerase is called polymerizing.
 6. Replication forks –

• LEADING STRAND
- The strand of nascent DNA
- It is synthesized in the same direction as the growing replication fork.
- The synthesis of leading strand is continuous.

• LAGGING STRAND
- is the strand of new DNA whose direction is opposite to the direction of the growing
replication fork
- Replication of the lagging strand is more complicated
• It is synthesized in short, separated segments, which are then joined together by DNA ligase
to form a continuous DNA strand.

• OKAZAKI FRAGMENT
- Short pieces synthesized DNA on the lagging strand.

*Replication can be unidirectional or bi directional.

7. Proof reading –

• Errors made during the replication may lead to mutation

• The DNA polymerase I and polymerase III act as proof readers of the
newly formed DNA
• They move along the new synthesized DNA, read mistakes formed
due to improper base pairing

8. Removal of RNA primer and completion of DNA strand

• The RNA primer is removed by DNA polymerase I which synthesizes
a short segment of complimentary DNA to seal the gap
• Polymerase I remove the primer one nucleotide at a time and replace it
with complimentary deoxyribonucleotide.
 9. Joining of fragments –

• Fragments are joined by DNA ligase

• A phosphodiesterase bond if formed between 3’ OH end of the
growing strand and 5’ end of the okazaki fragment.
DNA replication animations
https://www.youtube.com/watch?v=RLhwrFEtDEs

https://www.youtube.com/watch?v=3jslVQDGkLU
Protein Synthesis
https://www.youtube.com/watch?v=8wAwLwJAGHs

https://www.youtube.com/watch?v=bKIpDtJdK8Q