DNA Replication

Learning Objectives
 Explain the role of DNA polymerase in copying DNA.
 Compare DNA replication in prokaryotic cells and in
eukaryotic cells.
Review of DNA Structure

nitrogenous
bases
sugar-phosphate
backbone

double helix
Copying DNA

replication fork

DNA polymerase

Direction Direction
of replication of replication

new nucleotides
being added
 DNA polymerase joins nucleotides to synthesize a new, complementary strand of
DNA.
 the role of DNA polymerase: Besides producing the sugar-phosphate bonds that
join nucleotides together, DNA polymerase also “proofreads” each new DNA strand
so that each molecule is a near-perfect copy of the original.

 How is DNA unzipped at the replication forks? Hydrogen bonds are broken.

 What are the two roles of DNA polymerase in replication?

DNA polymerase joins individual nucleotides to produce a new strand of DNA and
proofreads the new strand.
DNA Replication

The blue strand represents the

original DNA strand.

The orange strand represents the

new DNA strand.

Is a new strand identical to its original

template strand?
No, they are complementary, not identical.
 DNA Replication
• Since the two strands of DNA are complementary each strand acts as a template for building
a new strand in replication.
• In DNA replication
 The parent molecule unwinds, and two new daughter strands are built based on base-pairing rules

DNA replication is semiconservative

Each of the two new daughter molecules will have one old strand, derived from the
parent molecule, and one newly made strand
 Okazaki
Leading & Lagging strands

Limits of DNA polymerase III

 can only build onto 3 end of
an existing DNA strand 5


rag ments
ki f
Okaza 5
3 5 5 3
3
5 Lagging strand
3
ligase
growing 3
replication fork
5
Leading strand

Lagging strand

3 5

3
DNA polymerase III
 Okazaki fragments
 joined by ligase Leading strand
 “spot welder” enzyme  continuous synthesis
 Summary
Initiation: DNA replication begins at specific locations called origins of
replication. Helicase unwinds the double helix, exposing the individual strands.
Single-strand binding proteins (SSBs) prevent the separated strands from
rejoining. Primase synthesizes short RNA primers that provide a starting point for
DNA polymerase to add complementary DNA nucleotides

Elongation: Two DNA polymerases—one for each strand—extend the nascent

DNA chains. Leading-strand synthesis is continuous, following the template
strand in the 5' to 3' direction. Lagging-strand synthesis occurs discontinuously
due to the antiparallel nature of the DNA strands, resulting in Okazaki fragments.
Each Okazaki fragment is extended in the 5' to 3' direction and requires post-
synthetic processing

Termination: Replication continues until the two replication forks meet and join
the newly synthesized DNA strands. Enzymes such as DNA ligase seal the gaps
between Okazaki fragments, creating full-length DNA strands
Telomeres
• Telomeres: the tips of eukaryotic chromosomes
• The enzyme telomerase adds short, repeated DNA
sequences to telomeres as the chromosomes are replicated.

Telomeres
The tips, or telomeres, of a DNA molecule are difficult to replicate. Cells use a
special enzyme called telomerase to add short repeated DNA sequences to
telomeres as chromosomes are replicated.

In the micrograph, the red structures are human chromosomes and the telomeres
are stained white, and appear in photograph as yellowish orange.
Prokaryotic DNA Replication

new DNA

replication fork

replication fork

unreplicated DNA
 Unlike eukaryotic DNA, the DNA of prokaryotic organisms
exists as a single loop. Explain that replication in most
prokaryotic cells begins at a single starting point and
proceeds in two directions until the entire chromosome is
copied.
Eukaryotic DNA Replication

Unreplicated DNA

Replication
forks

New DNA
 The process students looked at earlier in the presentation
was for replication eukaryotic DNA replication. Remind
students that eukaryotic chromosomes are linear.

 Where is the origin of replication?

in the center of each newly forming strand

 In eukaryotic cells replication may begin at dozens or

even hundreds of places on the DNA molecule,
proceeding in both directions until each chromosome is
completely copied.