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AP Bio Presentation - DNA Replication

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19 views20 pages

AP Bio Presentation - DNA Replication

Uploaded by

henryharkjin
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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DNA Replication

Eddie Kim
Why replicate DNA?
In multicellular organisms, cells divide for (1) growth and (2) reproduction.
• Growth is made possible by mitosis – a process in which a cell makes an identical
copy of itself.
• Reproduction is made possible by meiosis – a process in which one cell divides into
four non-identical gametes (eggs or sperm), each with only half the DNA.
• Single-celled organisms reproduce by simply copying themselves through mitosis.
For either mitosis or meiosis to occur, the DNA of the starting cell must be
replicated during interphase.
First Second
Parent cell replication replication

DNA Replication
(a) Conservative
Model: model

Semiconservative
(b) Semiconserva-
tive model

(c) Dispersive
model
New strand Template strand
5 end 3 end 5 end 3 end

Sugar A T A T
Phosphate Base

C G C G

G C G C

DNA polymerase
3 end A T A
T

C Pyrophosphate 3 end C

Nucleoside
triphosphate 5 end 5 end
DNA replication: overview

1) The replication process begins at an origin of


replication: a short “start here” sequence where
the multi-enzyme “replication complex” attaches to
the DNA.

2) Double-stranded DNA is pulled apart; each strand is


used to make a new complementary strand.

3) Replication proceeds in both directions from the


origin, creating a replication bubble where there
are now two identical sets of double-stranded DNA.

4) The moving ends of each bubble – where the


double-stranded DNA is getting pulled apart &
copied – are called the replication forks.
DNA replication: overview

Prokaryotes: One origin of replication on a circular DNA.


Opposite replication forks eventually meet, resulting in two
separate double-stranded DNAs.

Eukaryotes: Multiple origins of replication on each non-


circular DNA double helix. The multiple replication bubbles
eventually connect, resulting in two separate double-
stranded DNAs.
Replication enzymes to know

The DNA replication complex: a DNA-


making team that’s made up of many
individual enzymes working together on
their individual tasks.

You need to know only four major enzymes:


1) Helicase
2) Primase
3) DNA polymerases (collectively)
4) DNA ligase
Step 1: “Unzip” the DNA

Helicase separates the two parental strands


of the DNA double-helix so that they can be
replicated.
Informational Interlude
DNA polymerase
• The job: Build new a DNA strand by
1) Helping lone nucleotides base-
pair with nucleotides on the
parent strand, and
2) Catalyzing phosphodiester
bonds between incoming
nucleotides so they’re
connected to each other.
• The limitation: Can attach a new
nucleotide to the end of an existing
strand. Cannot bring in & bond
nucleotides when the “daughter
strand” doesn’t exist at all yet.
Step 2: Start adding primers

To compensate for this limitation of DNA


polymerase, Primase synthesizes a short
primer.
• Primer: A short stretch of RNA base-
paired to the DNA parental strand. It
serves as a starting block for DNA
polymerase to build a new complimentary
strand. RNA primers are replaced with
DNA later in the replication process.
Step 3: Build new complementary strands

???

parent
strand

???
Daughter strands are ALWAYS built 5’  3’ direction!

Whether in DNA replication or RNA


5’ end

transcription, new nucleotides are ALWAYS


added to the 3’ end of the growing strand
because bond formation is “fueled” by the
removal of two 5’ phosphates from the
incoming nucleotide.
* the covalent bond is called
“phosphodiester linkage”
3’ end

5’

3’
Consequences of 5’  3’ construction

• Double-stranded DNA is antiparallel


• If you follow one daughter strand from the origin of replication to the replication fork,
it’s oriented 5’  3. This strand can be built continuously. This daughter strand is the
leading strand.
Consequences of 5’  3’ construction

If you follow one daughter strand from the origin


of replication to the replication fork, it’s oriented
5’  3.
• This strand can be built continuously.

• It’s called the leading strand.

If you follow the other daughter strand from the


origin of replication to the replication fork, it’s
oriented 3’  5’.
• This strand CANNOT be built continuously.
It’s built in pieces called Okazaki
fragments.
Consequences of 5’  3’ construction
Step 3: Build new complementary strands

1) Primase makes primers


Step 3: Build new complimentary strands

1) Primase makes primers

2) A DNA polymerase III elongates the


new strands by adding DNA nucleotides
Step 4: Replace primers

1) Primase makes primers


2) A DNA polymerase III elongates the new
strands by adding DNA nucleotides
3) Once the fragments of the new strands touch,
another DNA polymerase I cuts out
the RNA primers and replaces them with
DNA nucleotides
Step 5: Join the Okazaki fragments

1) Primase makes primers


2) A DNA polymerase III elongates the new
strands by adding DNA nucleotides
3) Once the fragments of the new strands touch,
another DNA polymerase I cuts out the RNA
primers and replaces them with DNA
nucleotides.

4)Ligase joins together the remaining


adjacent nucleotides where the primers have
been replaced.
Assembling the Okazaki fragments

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