DIFFERENCE B/W PROKARYOTIC & EUKARYOTIC DNA
REPLICATION
• The circ.DNA molecules of the PK cell undergoes
replication in the cytoplasm itself which can
undergo Theta- model of replication
• The Lin. DNA molecule of the EK cell undergoes
replication inside the nucleoplasm during S-phase of
Interphase.
• During replication the 2 strands of DNA separate &
act as template for construction of a new strand on
them, as a result 2 DNA molecules are produced
• During replication free nucleotides which are
activated by Phosphorylase enzyme into
Triphosphate form come to the proximity of the old
parent strand according to Pairing rule & get paired
up
• The pairing is very fast in PK(2000 bps /sec) but slow
in EK(only 100-200 bps/sec)
• Replication starts from a special sequence called ori
(200-250 bps) inside the DNA molecule.
• Prokaryotic DNA has only 1 ori where replication
starts
• Whereas Eukaryotic DNA has many ori where
replication starts simultaneously or with small time
gaps.
• Later all the DNA fragments get joined by Ligases
MECHANISM OF REPLICATION IN GENERAL
• Step 1: Phosphorylation of true nucleotides in
cytoplasm into triphosphate
• This process is also called as activation /excitation
of nucleotides after which they enter the nucleus.
• Step 2: From the ori, the 2 strands of DNA starts
unwinding
• Step 3: The activated nucleotide gets arranged on the
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free strands according to pairing rule.
• The pairing rule has 5 postulates
• i) A pairs up with U or T
• ii) G pairs up with C
• iii) T pairs up with A
• iv) C pairs up with G
• v) Addition starts from 3' end of Old strand &
proceeds towards 5' end always ( In other words it
starts from 5' end of new strand & proceeds towards
3' end)
• Step 4: As we consider a Y-shaped replication fork ,
on 1 of the strands construction of new strand
becomes highly difficult bcoz of the 5th rule i.e on “A”
while on strand “B” construction of new strand is
very easy
• On strand A, the new strand is formed in fragments
after a small distance of unwinding is accomplished
& these goes on till the unwinding is completed.
• On strand A, for the formation of each Okazaki
fragment , a RNA primer has to be constructed first.
• While on strand B , RNA primer has to be constructed
only once.
• Step 5: Later on the RNA primers are detached & new
DNA nucleotides are substituted & the Okazaki
fragments are joined to form the complete strand
• Here the new strand formed on A is called
discontinuous strand & that formed on B is called
continuous strand
• Step 6: At the end of replication a process called
proof reading takes place which is conducted in 3' to
5' direction of new strand for making it perfect. This
is also called as major repair.
• TABLE OF ENZYMES INVOLVED IN DNA REPLICATION:
To be discussed in class