Scribd
Upload
14 views27 pages

BIO1140 Topic 7 Replication W2021

Uploaded by

Yunsu Lee
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
14 views27 pages

BIO1140 Topic 7 Replication W2021

Uploaded by

Yunsu Lee
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
You are on page 1/ 27

BIO1140

Topic 7 – Replication
Learning Outcomes
By the end of this topic you should be able to:
• understand, describe and draw how DNA replication occurs at the
molecular level, from initiation through to completion, this includes:
• Describing the importance of replication origins, key differences between
prokaryotes and eukaryotes, and the process by which initiation of replication
occurs at these origins
• Naming the various proteins involved in DNA replication, describing their
function, and associating them with the specific stage(s) of replication they
participate it
• Distinguishing the template strand from the newly synthesized daughter strand
during replication
• Appreciating the asymmetrical nature of the replication fork, describing the
underlying molecular events that cause it to be asymmetrical, distinguishing the
leading (continuous) from the lagging (discontinuous) strand, and discussing the
concept of Okazaki fragments
• Describing the importance of telomeres and how they are replicated
Be sure you understand the structure of DNA well before continuing
• review the HHMI chemical structure of DNA video as needed
The relationship between the structure and function of DNA became apparent
once its structure was determined
• Watson and Crick quickly predicted the semiconservative model of DNA
replication (Meselson and Sthal later confirmed it experimentally)
i.e. - during DNA replication, the parent molecule unwinds, each strand acts as a template,
and two new daughter strands are built based on base-pairing rules

Campbell Biology - Figure 16.9


Replication
Nature Scitable – DNA replication

3 key phases to consider:


1) Initiation
2) Elongation
3) Termination

At the beginning of replication, DNA is unzipped by helicase


(yellow). Then primase (red) creates the first few base pairs
that DNA polymerase (blue) will recognize, before it begins
replication.
Replication - Initiation

• Replication begins at sites called


origins of replication, where two
DNA strands are separated, opening a
replication “bubble”
• A bacterial chromosome will typically
have one origin of replication
Campbell Biology - Figure 16.12
Replication - Initiation

• A eukaryotic chromosome may have


hundreds or thousands of origins of
replication
• Replication proceeds in both directions
from each origin, until the entire molecule
is copied

Campbell Biology - Figure 16.12


At the end of each replication
bubble is a replication fork, a
Y-shaped region where new
DNA strands are elongating
Helicases are enzymes that untwist the
DNA double helix at the replication forks.
Topoisomerase is a family of enzymes
which correct the “overwinding” ahead of
replication forks by breaking, swiveling,
and rejoining DNA strands.
Single-strand binding proteins bind to
and stabilize single-stranded DNA in
its linear form.
Primase can start an RNA chain de
novo and add RNA nucleotides one
at a time using parental DNA as a
template.
• The resulting primer is short (5–10
nucleotides), and the 3 end serves
as starting point for the synthesis
of DNA by DNA polymerase
Replication Initiation
Summary

Campbell Biology - Figure 16.13


DNA Polymerase DNA polymerase has 2
requirements:
• DNA synthesis is carried out by a • Free deoxynucleotide triphosphates
family of enzymes known as DNA (dGTP, dATP, dCTP and dTTP)
polymerases • A template strand of DNA with a
• They add nucleotides in the 5’-3’ short double stranded primer region
direction with an exposed 3’-OH
Replication - Elongation
• DNA polymerase catalyzes the
formation of phosphodiester bonds,
elongating the new DNA strand
• As each new nucleotide is added to
the growing DNA strand, it loses two
phosphate groups as a molecule of
pyrophosphate
• The rate of elongation is about:
• 500 nucleotides per second in bacteria
• 50 nucleotides per second in human
cells

Campbell Biology - Figure 16.14


Elongation

The antiparallel structure of the double helix


affects replication
• DNA polymerases add nucleotides only to
the free 3 end of a growing strand
• Therefore, a new DNA strand can only
elongate in the 5 to 3 direction

Campbell Biology - Figure 16.15


• Along one template strand of DNA, DNA
polymerase III synthesizes the leading
strand continuously, moving toward the
replication fork
Campbell Biology - Figure 16.15
Synthesis of the Lagging Strand
• To elongate the other new strand, the lagging
strand, DNA polymerase III must work in the
direction away from replication fork
• The lagging strand is synthesized
discontinuously as series of segments called
Okazaki fragments

Campbell Biology - Figure 16.16


Synthesis of the Lagging Strand

• Synthesis of each Okazaki fragment begins


with an RNA primer generated by primase
• DNA polymerase III then completes the
Okazaki fragment, stopping and dissociating
at the next RNA primer it encounters
Campbell Biology - Figure 16.17
Synthesis of the Lagging Strand

• Upon completion of the next Okazaki


fragment, the RNA primers are removed and
replaced with DNA by DNA polymerase I
• DNA ligase seals the Okazaki fragments
generating a single continuous DNA strand

Campbell Biology - Figure 16.17


Synthesis of the
Lagging Strand
Summary

Campbell Biology - Figure 16.17


Replication –
Summary of The Players

Campbell Biology - Table 16.1


Replication Elongation
Summary

Campbell Biology - Figure 16.17


Replication Complex

Proteins participating in
DNA replication form a
large complex known as the
DNA replication machine
or the replisome which
significantly increases the
efficiency of the process.

Campbell Biology - Figure 16.18


Replication - Termination

Limitations of DNA polymerase create problems


for linear DNA of eukaryotic chromosomes
• The usual replication machinery provides no
way to complete the 5 ends after the last
RNA primer is removed
• Therefore, repeated rounds of replication
result in shortening of DNA molecules at
the ends
• Left unchecked this can lead to loss of
genes from a chromosome
• This is not a problem for prokaryotes, as most
have circular chromosomes

Campbell Biology - Figure 16.20


• Eukaryotic chromosomal DNA molecules have special nucleotide sequences at
their ends called telomeres
• Telomere shortening has been associated with aging

Replication origin

Telomere

Campbell Biology - Figure 16.21 and 16.22


• Telomerase is a protein-RNA
complex which recognizes the end of
a telomere sequence and elongates
it (5’-3’) using its own built in RNA
template
• DNA polymerase then completes the
other strand preventing excessive
shortening and loss of genes from
the chromosome

You might also like