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DNA:
The Genetic Material
What is DNA?
Deoxyribonucleic Acid
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The DNA Structure
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• Genetic information in cells is embedded in
the nucleic acids DNA and RNA.
• Deoxyribonucleic acid, DNA, carries
the genetic blueprint for the cell while
ribonucleic acid, RNA, is an inter-
mediary molecule that converts this
blueprint into defined amino acid
sequences in proteins.
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The monomers of nucleic acids are called
nucleotides, consequently, DNA and RNA
are polynucleotides.
A nucleotide has three components:
1. a pentose sugar (either ribose in
RNA or deoxyribose in DNA),
2. a nitrogen base, and
3. a molecule of phosphate, PO43-.
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• The nucleotides are covalently bonded
by phosphate between the 3′- (3 prime)
carbon of one sugar and the 5′-carbon
of the next sugar.
• The phosphate linkage is called a
phosphodiester bond because the
phosphate connects two sugar
molecules by an ester linkage
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❖ In the genome of cells, DNA is double-
stranded.
❖ The strands are held together by
hydrogen bonds that form between the
bases in one strand and those of the
other strand. When located adjacent to
one another, purine and pyrimidine
bases can form hydrogen bonds
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Hydrogen bonding is most thermodynamically
stable when guanine (G) bonds with cytosine
(C) and adenine (A) bonds with thymine (T).
Specific base pairing, A with T and G with C,
ensures that the two strands of DNA are
complementary in base sequence; that is,
wherever a G is found in one strand, a C is found
in the other, and wherever a T is present in one
strand, its complementary strand contains an A.
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The molecular processes of genetic information flow can be divided
into three stages:
1.Replication. During replication, the DNA double helix
is duplicated, producing two copies. Replication is
carried out by an enzyme called DNA polymerase.
2. Transcription. The transfer of genetic information
from DNA to RNA is called transcription. Transcription is
carried out by an enzyme called RNA polymerase.
3. Translation. Synthesis of a protein, using the genetic
information in mRNA, is called translation.
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DNA Replication
Transmission of Genetic Information
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Initiation of DNA Synthesis
Before DNA polymerase can synthesize new DNA,
the double helix must be unwound to expose the
template strands.
The zone of unwound DNA where replication
occurs is called the replication fork. The enzyme
DNA helicase unwinds the double helix, using
energy from ATP, and exposes a short single-
stranded region
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Initiation of DNA Synthesis
Helicase moves along the DNA and separates the
strands just in advance of the replication fork. The
single-stranded region is immediately covered with
copies of single- strand binding protein to stabilize
the single-stranded DNA and prevent the double
helix from re-forming.
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Initiation of DNA Synthesis
Next, primase and then DNA polymerase
enzymes are loaded onto the DNA behind the
helicases. Initiation of DNA replication then begins
on the two single strands.
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Replication always proceeds from 5′ to 3′ (always
adding a new nucleotide to the 3′-OH of the
growing chain).
On the strand growing from the 5′-PO42- to the 3′-
OH, called the leading strand, DNA synthesis
occurs continuously because there is always a
free 3′-OH at the replication fork to which a new
nucleotide can be added.
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❖ But on the opposite strand, called the lagging
strand, DNA synthesis occurs discontinuously
because there is no 3′-OH at the replication fork
to which a new nucleotide can attach.
❖ Therefore, on the lagging strand, RNA primers
must be synthesized by primase multiple times
to provide free 3′-OH groups.
❖ As a result, the lagging strand is made in short
segments, called Okazaki fragments, named
after their discoverer, Reiji Okazaki. These
lagging strand fragments are joined together
later to yield a continuous strand of DNA.
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Synthesis of the New DNA Strands
After synthesizing the RNA primer, primase is
replaced by another DNA Polymerase. This
enzyme is actually a complex of several proteins
including the polymerase core enzyme itself. Each
molecule of polymerase is held on the DNA by a
sliding clamp, which encircles and slides along the
single template strands of DNA.
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When the primer has been removed and replaced
with DNA, DNA Polymerase is released.
The very last phosphodiester bond is made by an
enzyme called DNA ligase. This enzyme seals
nicks in DNAs that have an adjacent 5′-PO42- and
3′-OH and along with DNA Polymerase, it also
participates in DNA repair.
DNA ligase is also important for sealing genetically
manipulated DNA during molecular cloning.
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the Replisome
Instead, replication proteins aggregate to form a
large replication complex called the replisome.
The lagging strand of DNA actually loops out to
allow the replisome to move smoothly along both
strands, and the replisome literally pulls the DNA
template through it as replication occurs.
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the replisome contains several key
replication proteins:
(1) DNA gyrase, which removes supercoils;
(2) DNA helicase and primase (the primo-
some), which unwind and prime the DNA;
and
(3) single-strand binding protein, which
prevents the separated template strands
from re-forming a double helix
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Fidelity of DNA replication: Proofreading
DNA replicates with a remarkably low error rate.
Nevertheless, when errors do occur, a mechanism
exists to detect and correct them. Errors in DNA
replication introduce mutations, changes in DNA
sequence. Mutation rates in cells are remarkably
low, between 10−8 and 10−11 errors per base pair
inserted. This accuracy is possible partly because
DNA polymerases get two chances to incorporate
the correct base at a given site.
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Termination of replication
Eventually the process of DNA replication is
finished. How does the replisome know when to
stop?
On the opposite side of the circular chromosome
from the origin is a site called the terminus of
replication.
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The two replication forks collide as the new circles
of DNA are completed. In the terminus region are
several DNA sequences called Ter sites that are
recognized by a protein called Tus, whose function
is to block progress of the replication forks.
When replication of the circular chromosome is
complete, the two circular molecules are linked
together, much like the links of a chain.
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Then, they are unlinked by another enzyme,
topoisomerase IV. Obviously, it is critical that, after
DNA replication, the DNA is partitioned so that each
daughter cell receives a copy of the chromosome.
This process may be assisted by the important cell
division protein FtsZ, which helps orchestrate
several key events of cell division.
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Any questions?
This presentation is already available for download in our google classroom.
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Assignment:
Download the DNA Model activity sheet from our
google class library. Read the instructions and
perform the tasks indicated in the activity sheet.
Date of Submission: August 1, 2019
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