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DNA replication is the process in which a cell’s entire DNA is copied, or replicated. The
identification of the structure of DNA suggested that each strand of the double helix would serve
as a template for synthesis of a new strand. DNA replication process occurs during the Synthesis
(S) phase of the eukaryotic cell cycle. As each DNA strand has the same genetic information,
both strands of the double helix can serve as templates for the reproduction of a complementary
new strand. The two resulting double helices, which each contain one "old" strand and one "new"
strand of DNA, are identical to the initial double helix. DNA replication is said to be semi-
conservative because of this process of replication, where the resulting double helix is composed
of both an old strand and a new strand.
DNA replication occurs when the DNA strands “unzip”, and the original strands of DNA serve
as a template for new nucleotides to join and form a new strand. The replication fork is obvious
at the point of separation of the double helix.
The semi-conservative mechanism of replication was one of three models originally proposed for
DNA replication:
1. Semiconservative replication would produce two copies that each contained one of the
original strands and one new strand.
2. Conservative replication would leave the two original template DNA strands together in a
double helix, with the new DNA composed entirely of two new strands.
3. Dispersive replication would produce two copies of the DNA, both containing a mixture
of old and new material.
Of these three models, the semi-conservative model seemed most reasonable since it would
allow each new daughter strand to remain associated with its template strand. The
semiconservative model was confirmed by the Meselson-Stahl experiment.
DNA replication is a semi-conservative process. Half of the parent DNA molecule is conserved
in each of the two daughter DNA molecules.
In 1958, Matthew Meselson and Franklin Stahl identified evidence that supported the hypothesis
that DNA replication was semiconservative. The experiment took advantage of the fact that
nitrogen is a major constituent of DNA, and that two forms of nitrogen are available: 14N and a
heavier isotope, 15N. The DNA of cells grown in 15N medium have a higher density than cells
grown in normal 14N medium. The experiment was performed as follows:
DNA replication begins as an enzyme, DNA helicase, breaks the hydrogen bonds holding the
two strands together and forms a replication fork. The resulting structure has two branching
strands of DNA backbone with exposed bases. These exposed bases allow the DNA to be “read”
by another enzyme, DNA polymerase, which then builds the complementary DNA strand. As
DNA helicase continues to open the double helix, the replication fork grows.
5' → 3
The two new strands of DNA are “built” in opposite directions, through either a leading strand
or a lagging strand. The leading strand is the DNA strand that DNA polymerase constructs in
the 5' → 3' direction. This strand of DNA is made in a continuous manner, moving as the
replication fork grows. The lagging strand is the DNA strand at the opposite side of the
replication fork from the leading strand. It goes in the opposite direction, from 3' to 5'. DNA
polymerase cannot build a strand in the 3' → 5' direction. Thus, this "lagging” strand is
synthesized in short segments known as Okazaki fragments. On the lagging strand, an enzyme
known as primase builds a short RNA primer. DNA polymerase is then able to use the free 3'-
OH group on the RNA primer to make DNA in the 5' → 3' direction. The RNA fragments are
then degraded and new DNA nucleotides are added to fill the gaps where the RNA was present.
Another enzyme, DNA ligase, is then able to attach (ligate) the DNA nucleotides together,
completing the synthesis of the lagging strand (Figure below).
DNA replication. The two DNA strands are opened by helicase. The strands are held open by a
single strand of binding proteins, preventing premature reannealing. Topoisomerase solves the
problem caused by tension generated by winding/unwinding of DNA. This enzyme wraps around
DNA and makes a cut permitting the helix to spin and relax. Once DNA is relaxed,
topoisomerase reconnects broken strands. DNA primase synthesizes a short RNA primer which
initiates the Okazaki fragment. Okazaki fragments are attached by DNA ligase.
Many replication forks develop along a chromosome. This process continues until the replication
forks meet, and the all of the DNA in a chromosome has been copied. Each new strand that has
formed is complementary to the strand used as the template. Each resulting DNA molecule is
identical to the original DNA molecule. During prophase of mitosis or prophase I of meiosis,
these molecules of DNA condense into a chromosome made of two identical "sister" chromatids.
This process ensures that cells that result from cell division have identical sets of genetic
material, and that the DNA is an exact copy of the parent cell’s DNA.
Vocabulary
DNA helicase: The enzyme that breaks the hydrogen bonds holding the two DNA strands
together during DNA replication.
DNA ligase: Enzyme that can attach together (ligates) strands of DNA with double strand
breaks.
DNA polymerase: The enzyme that builds a new DNA strand during DNA replication.
DNA replication: The process of copying DNA prior to cell division (eukaryotes) or
reproduction (prokaryotes).
lagging strand: The DNA strand at the opposite side of the replication fork from the
leading strand.
leading strand: The DNA strand that DNA polymerase constructs in the 5' → 3'
direction.
Okazaki fragments: Short fragments of DNA that comprise the lagging strand.
primase: An enzyme that builds a short RNA primer on the lagging strand during DNA
replication.
replication fork: Site where DNA helices unwinds the DNA, allowing DNA replication
to proceed.
semi-conservative: DNA replication process where half of the parent DNA molecule is
conserved in each of the two daughter DNA molecules.
Summary
DNA replication is the semi-conservative process by which a cell’s entire DNA is copied,
or replicated.
During DNA replication, the two new strands of DNA are “built” in opposite directions,
starting at replication forks.
Review
Read these passages from the text and answer the questions that follow.
DNA
DNA is the genetic material in your cells. It was passed on to you from your parents and
determines your characteristics. The discovery that DNA is the genetic material was another
important milestone in molecular biology.
Based on his observations, Griffith deduced that something in the killed S-strain was transferred
to the previously harmless R-strain, making the R-strain deadly. What was that something? What
type of substance could change the characteristics of the organism that received it?
The conclusion that DNA is the genetic material was not widely accepted at first. It had to be
confirmed by other research. In the 1950s, Alfred Hershey and Martha Chase did experiments
with viruses and bacteria. Viruses are not cells. They are basically DNA inside a protein coat. To
reproduce, a virus must insert its own genetic material into a cell (such as a bacterium). Then it
uses the cell’s machinery to make more viruses. The researchers used different radioactive
elements to label the DNA and proteins in viruses. This allowed them to identify which molecule
the viruses inserted into bacteria. DNA was the molecule they identified. This confirmed that
DNA is the genetic material.
Questions
2. Why did the rough strain and heat-killed smooth strain kill the mice?
1. Which of the following statements concerning DNA is correct? (1) DNA contains
instructions for all the proteins your body makes. (2) The shape of DNA is a double helix.
(3) The central dogma of molecular biology states RNA → DNA → Protein.
1. 1 only
2. 1 and 2
3. 2 and 3
4. 1, 2, and 3
5. Differences between DNA and RNA include which of the following? (1) RNA consists
of one nucleotide chain. (2) RNA contains the nitrogen base uracil instead of thymine. (3)
RNA contains the sugar ribose instead of deoxyribose.
1. 1, 2, and 3
2. 1 and 2
3. 2 and 3
4. 2 only
Lesson 7.1: Vocabulary I
Name___________________ Class______________ Date________
Definitions
_____ 2. found that there exists a substance that could change the characteristics of another
organism
_____ 10. copies the genetic instructions from DNA in the nucleus, and carries the instructions
to the cytoplasm
Terms
b. Chargaff's rules
c. DNA replication
d. double helix
e. Franklin
f. Griffith
i. nucleotide
1. In DNA, A always pairs with ____________, and G always pairs with ____________.
3. Griffith showed that a substance could be transferred to harmless bacteria and make them
____________.
6. The amount of A equals the amount of T and the amount of G equals the amount of C is
known as ____________ rules.
7. ____________ RNA copies the genetic instructions from DNA in the nucleus, and carries
them to the cytoplasm.
_____ 1. The process in which cells make proteins is called protein expression.
_____ 2. Transcription takes place in three steps: initiation, elongation, and termination.
_____ 5. UAG, UGA, AGU, and UAA are the four stop codons
_____ 7. Initiation of transcription occurs when the enzyme, DNA polymerase, binds to the
promoter of a gene.
_____ 8. All known living organisms, except some species of primitive bacteria, have the same
genetic code.
_____ 11. Many proteins are modified in the Golgi apparatus after translation.
_____ 12. During translation, rRNA brings the amino acids into the ribosome.
How is the information in a gene encoded? The answer is the genetic code. The genetic code
consists of the sequence of nitrogen bases — A, C, G, T (or U) — in a polynucleotide chain. The
four bases make up the “letters” of the genetic code. The letters are combined in groups of three
to form code “words,” called codons. Each codon stands for (encodes) one amino acid, unless it
codes for a start or stop signal.
There are 20 common amino acids in proteins. There are 64 possible codons, more than enough
to code for the 20 amino acids. The genetic code is shown in the FlexBook.
As shown in the Genetic Code figure (see Figure 7.8 in the FlexBook), the codon AUG codes
for the amino acid methionine. This codon is also the start codon that begins translation. The
start codon establishes the reading frame of mRNA. The reading frame is the way the letters are
divided into codons. After the AUG start codon, the next three letters are read as the second
codon. The next three letters after that are read as the third codon, and so on. The mRNA
molecule is read, codon by codon, until a stop codon is reached. UAG, UGA, and UAA are all
stop codons. They do not code for any amino acids.
The genetic code is universal. All known living things have the same genetic code. This
shows that all organisms share a common evolutionary history.
The genetic code is unambiguous. Each codon codes for just one amino acid (or start or
stop). What might happen if codons encoded more than one amino acid?
The genetic code is redundant. Most amino acids are encoded by more than one codon.
What might be an advantage of having more than one codon for the same amino acid?
Questions
5. What might happen if codons encoded more than one amino acid?
Lesson 7.2: Multiple Choice
Name___________________ Class______________ Date________
3. Which of the following terms is most closely associated with a tRNA molecule?
1. codon
2. anticodon
3. transcription
4. ribosome
Definitions
_____ 5. contains an anticodon that is complementary to the codon for an amino acid
_____ 11. the way the groups of three bases are divided into codons
_____ 12. a region of a gene where RNA polymerase binds
Terms
a. codon
b. exons
c. genetic code
d. introns
e. mRNA
f. promoter
g. protein synthesis
h. reading frame
i. splicing
j. tRNA
k. transcription
l. translation
6. ____________ is the second part of the central dogma of molecular biology: RNA → Protein.
7. Polyadenylation adds a “tail” of ____________ to the mRNA.
8. AUG is the start codon and it codes for the amino acid ____________.
10. The mRNA molecule is read, codon by codon, until a ____________ codon is reached.
_____ 5. Neutralor silent mutations can have a significant effect on the organism.
_____ 6. A deletion or insertion of one or more nucleotides may result in a frameshift mutation.
_____ 9. The cell does not have the capability to repair damaged DNA.
Read these passages from the text and answer the questions that follow.
Beneficial Mutations
Some mutations have a positive effect on the organism in which they occur. They are called
beneficial mutations. They lead to new versions of proteins that help organisms adapt to changes
in their environment. Beneficial mutations are essential for evolution to occur. They increase an
organism’s changes of surviving or reproducing, so they are likely to become more common
over time. There are several well-known examples of beneficial mutations. Here are just two:
1. Mutations in many bacteria that allow them to survive in the presence of antibiotic drugs.
The mutations lead to antibiotic-resistant strains of bacteria.
2. A unique mutation is found in people in a small town in Italy. The mutation protects them
from developing atherosclerosis, which is the dangerous buildup of fatty materials in
blood vessels. The individual in which the mutation first appeared has even been
identified.
Harmful Mutations
Imagine making a random change in a complicated machine such as a car engine. The chance
that the random change would improve the functioning of the car is very small. The change is far
more likely to result in a car that does not run well or perhaps does not run at all. By the same
token, any random change in a gene's DNA is likely to result in a protein that does not function
normally or may not function at all. Such mutations are likely to be harmful. Harmful mutations
may cause genetic disorders or cancer.
Questions
4. A frameshift mutation
1. changes the reading frame of the base sequence.
2. can be due to a translocation between two chromosomes.
3. may not have an effect on how the codons in mRNA are read.
4. all of the above.
5. Beneficial mutations
1. may only cause mild genetic disorders.
2. are silent mutations, which code for the same amino acid.
3. help organisms adapt to changes in their environment.
4. are only caused by beneficial mutagens, like barbecuing and tanning.
6. A missense mutation
1. results in a premature stop codon.
2. codes for a different amino acid.
3. codes for the same amino acid.
4. results in a frameshift mutation.
Definitions
_____ 1. a deletion or insertion of one or more nucleotides that changes the reading frame
_____ 12. generally caused by mutations in genes that regulate the cell cycle
Terms
a. cancer
b. chromosomal alteration
c. deletion
d. duplication
e. frameshift mutation
f. genetic disorder
g. germline mutations
h. insertion
i. inversion
j. mutagen
k. mutation
l. point mutation
m. somatic mutations
n. translocation
5. Mutations are essential for ____________ to occur because they increase genetic variation.
11. ____________ mutations have a positive effect on the organism in which they occur.