DNA Replication I

DNA replication I
Wednesday, June 26, 2024

7:46 PM
Genomes size ( C- value)

 C-value is the amount of DNA in the unreplicated gametic nucleus.
 Can be measured as either number of base pairs (bp) ot the mass of DNA in a haploid
chromosome.
 10*6 bp = 1 Mbp = 1 fg (femtogram) DNA. The actual number of base pairs or mass
of DNA in the cell of a diploid organism is therefore twice that shown here. For example, the
human euchromatic genome is 3.2 x 10*9 bp, the E.coli genome is about 4.6 x 10*6 bp
In bacteria 90% of the genome has a purpose for coding and regulatory sequences.
Whereas in the eukaryotic cells on 1% of the genomes codes for proteins. Most of the eukaryotic
genome codes for RNA which has other purposes.
Gene composition
Humans = 30,000-40,000
Gut bacterium= 4000
Gene = Basic Heredity and a sequence of nucleotides in DNA that encodes the synthesis of a gene
product.
During Gene expression the DNA is first copied into RNA that can be functional or a template for a
protein.
https://www.nature.com/scitable/topicpage/cells-can-replicate-their-dna-precisely-
6524830/
DNA replication
- Double-stranded DNA molecule is copied to produce two identical DNA molecules.
- During replication, each DNA strand acts as a template for the production of a new DNA
strand.
- A replication fork is formed, where the DNA unwinds and two new strands are synthesized.
- Fidelity is maintained by complementary base pairing: A pairs with T, and G pairs with C.
- New DNA can only be synthesized in a 5’ to 3’ direction.
 dNTPs consist of a deoxyribose sugar, a nitrogenous base (A, T, C, or G), and three
phosphate groups.
 DNA polymerase adds dNTPs to the growing DNA strand during replication.
 The enzyme pairs each dNTP with its complementary base on the template strand (A
pairs with T, G pairs with C).
 DNA synthesis starts of with the synthesis of short length of RNA, complementary to
the DNA strand being copied, to act as a primer.
 Helicase = An enzyme that uses ATP to move along the DNA strand to allow DNA helix to
allow for replication
 Replication fork = localised region of replication, where the two strands separate to allow for
the replication to take place.
 Template strand = original DNA strand that is being copied.
 DNA polymerase = synthesises DNA by joining nucleotides together using a DNA template as
a guide.
The two DNA strands :
 Leading strand = The newly synthesised strand from 5' to 3'
 Lagging strand = DNA synthesis has to take place in 5' to 3' only. However the DNA strands of
the DNA run in opposite directions antiparallel . This means lagging strand takes longer to
complete than the leading strand. The lagging strand is synthesized discontinuously in small
segments known as Okazaki fragments.
 DNA ligase seals the gaps between the Okazaki fragments, joining the fragments into a single
DNA molecule
Topoisomerase:

Sliding Clamp PCNA :
 Function is to hold polymerase enzyme tightly onto the DNA during replication. This means
polymerase can synthesise long stretches of DNA without falling off.
 During DNA replication and transcription, the DNA helix needs to be unwound. This
unwinding can create tension and cause the DNA ahead of the replication fork to become
supercoiled (over-twisted).
 Topoisomerases prevent or resolve these supercoils by cutting the DNA, allowing it to
unwind, and then resealing it.
 The origin is the site where DNA replication starts

 In E.coli, the origin is called oriC.
 In eukaryotes there are multiple origins of replication on each chromosome as the
eukaryotic genome is much larger than the prokaryotic one.
Semi-Conservative Replication: This result showed that DNA replicates in a semi-conservative way.
Each new DNA molecule consists of one strand from the original molecule (heavy or medium) and
one newly synthesized strand (light).
In prokaryotic cells DNA replication begin a one point known as the OriC.
From this point it begins replication in both directions (BI-DIRECTIONAL)
Replication takes place close to the replication fork.
REPLICON
 A unit of DNA where an individual act of replication occurs is called a replicon.
 It contains the control elements that are required for replication
 Contains an origin
 May also contain a terminus
A replicated region looks like an eye in the circle of DNA .
The structural difference in DNA in prokaryotic and eukaryotic DNA means DNA replication can be
different
Steps of DNA replication
1. DNA unwinds at the origin of replication.
2. Helicase opens up the DNA-forming replication forks; these

are extended in both directions.
3. Single-strand binding proteins coat the DNA around the

replication fork to prevent rewinding of the DNA.
4. Topoisomerase binds at the region ahead of the replication

fork to prevent supercoiling (over-winding).
5. Primase synthesizes RNA primers complementary to the DNA

strand.
6. DNA polymerase III starts adding nucleotides to the 3′-OH

(sugar) end of the primer.
7. Elongation of both the lagging and the leading strand

continues.
8. RNA primers are removed and gaps are filled with DNA by
DNA pol I.
9. The gaps between the DNA fragments are sealed by DNA

ligase.

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DNA replication I

Wednesday, June 26, 2024

Genomes size ( C- value)

 The origin is the site where DNA replication starts

1. DNA unwinds at the origin of replication.

2. Helicase opens up the DNA-forming replication forks; these

3. Single-strand binding proteins coat the DNA around the

4. Topoisomerase binds at the region ahead of the replication

5. Primase synthesizes RNA primers complementary to the DNA

6. DNA polymerase III starts adding nucleotides to the 3′-OH

7. Elongation of both the lagging and the leading strand

9. The gaps between the DNA fragments are sealed by DNA

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