Lesson 2.
DNA Replication
and Protein
Synthesis
General Biology 1/2
2
Science, Technology, Engineering, and Mathematics
With the enormous
number of living
organisms on the
planet, how does
cell have their own
copy of DNA?
2
Do cells produce
their own unique
version of their
DNA? Do they rely
on the copy of the
DNA from
preexisting cells?
3
In addition, how is DNA ‘converted’ into proteins
such as this green fluorescent protein from the
sea jelly Aequorea victoria?
4
How is it possible for
thousands of cells in cell
culture to have almost
identical genetic material?
5
Learning Competency
At the end of the lesson, you should be able to do the
following:
Diagram the steps in DNA replication and
protein synthesis (STEM_BIO11/12-IIIa-b-
5).
6
Learning Objectives
At the end of the lesson, you should be able to do the
following:
● Explain the process of DNA replication.
● Explain the processes involved in transcription
and translation.
7
Reproduction and DNA Replication
The
duplication
of the
genetic
material is a
key process
in the
reproduction
of living cells.
8
Reproduction and DNA Replication
GROWTH
AND
DEVELOPME
NT
DNA
REPLICATIO
N
WOUND ASEXUAL
HEALIN REPRODUCTI
G ON
9
Semi-conservative Nature of
Replication
The three major models of DNA replication 10
Semi-conservative Nature of
Replication
The Meselson-
Stahl experiment
proved the
semiconservative
nature of DNA
replication.
11
Mechanism of Replication
Each strand serves
as a template.
Resulting DNAs
have an old and a
new strand.
Replication is
through
complementary
base pairing.
Parental DNA is
not completely
conserved. 12
Mechanism of Replication
Replication bubble is the site where the bidirectional
and 5’-to-3’ DNA replication actively takes place. 13
Origin of Replication
14
Initiation of Replication
stabilization of
unwinding of the single-
the DNA stranded DNA
bidirection
relieves tension al
in the replication
unwinding DNA
15
Initiation of Replication
Priming is an essential step before DNA polymerase continues
with the elongation of the DNA strand.
16
Elongation in Replication
This is
replication
as seen in
one end of
the fork.
17
Elongation in Replication
Topoisomerase is
the eukaryotic
counterpart of
gyrase.
18
Elongation in Replication
Separate
polymerases act
on leading and
lagging strand.
19
Elongation in Replication
The polymerases
synthesize DNA in the
5’ to 3’ direction.
20
Elongation in Replication
Synthesis occurs
continuously in the
leading strand.
21
Elongation in Replication
Synthesis occurs in
fragments in the
lagging strand.
22
Elongation in Replication
Ligase joins the
Okazaki fragments
together.
23
Summary of Replication Proteins
Replication Function
Protein
Attaches to the origin of replication and creates an
Initiator protein
initial separation between the two DNA strands
Continuously unwinds the DNA at the replication
DNA helicase
fork
Single-strand
Bind to the single-stranded DNA to prevent them
DNA-binding
from reforming hydrogen bonds
proteins
DNA gyrase (or Moves ahead of the DNA helicase at the replication
topoisomerase fork and relieves the tension that builds up as the 24
Summary of Replication Proteins
Replication Function
Protein
Synthesize short ribonucleotide segments called
DNA primase RNA primers to provide a 3’-OH for the DNA
polymerase III
Synthesizes deoxynucleotide chains from the 3’-OH
DNA polymerase
end of the RNA primers and performs DNA
III
proofreading via its exonuclease activity
DNA polymerase Catalyzes the removal of the RNA primers and
I replaces them with DNA nucleotides
Seals the nicks between the Okazaki fragments 25
Termination of Replication
26
How do cells minimize the
errors that occur during
DNA replication?
27
DNA Repair
28
How can cells from different
species produce proteins
with identical amino acid
sequences?
29
Central Dogma of Molecular Biology
DNA as the
RNA as the proteins as the
repository of
transcript of genetic phenotypic
genetic
information determiners
information
30
Types of RNA Molecules
GENE TRANSLATION AMINO ACID
TRANSCRIPT MACHINERY CONVEYORS
31
Types of RNA Molecules
RNA Class Location in Cells Function
Messenger RNA Serves as the transcript for
Nucleus and cytoplasm
(mRNA) the amino acid sequence
Transfer RNA Carries amino acids to the
(tRNA) Cytoplasm ribosomes for protein
synthesis
Ribosomal RNA Structural and functional
Cytoplasm
(rRNA) component of ribosomes
Small nuclear RNA Processing of pre-mRNA to
Nucleus
(snRNA) mature mRNA
Small nucleolar Processing of rRNA for
Nucleus
RNA (snoRNA) ribosomes 32
The Process of Transcription
Transcription
factors bind to
the promoter
region.
33
The Process of Transcription
RNA polymerase
binds to the
promoter region
through the
sigma factor.
34
The Process of Transcription
RNA polymerase
unwinds the DNA.
35
The Process of Transcription
Polymerase
transcribes the
coding
sequence
36
The Process of Transcription
Terminator sequence is
encountered.
37
The Process of Transcription
Polymerase
and RNA
dissociate
from the
complex.
38
The Process of Transcription
The gene is
completely
transcribed.
39
Post-Transcriptional Modification
The resulting RNA undergoes 5’-methyl capping, intron splicing,
and addition of a poly-A tail. 40
What do you think are the
possible consequences if a
cell fails to perform one of
the chemical modifications
to the mRNA transcript?
41
The Genetic Code
Each codon
of the mRNA
corresponds
to an amino
acid.
42
The Genetic Code
43
The Translation Process
Ribosomal
subunits, mRNA,
and first tRNA
assemble.
44
The Translation Process
First tRNA
binds to the
P site (with
AUG codon
in mRNA).
45
The Translation Process
The next
tRNA enters
the A site.
46
The Translation Process
Peptide bond
forms
between the
amino acids. 47
The Translation Process
Amino acid
chain is now
at the A site
tRNA.
48
The Translation Process
The ribosome
moves to the 3’
direction.
49
The Translation Process
The tRNA at E
site leaves.
50
The Translation Process
The A site is
now
available for
the next
tRNA.
51
What do you think will
happen if an error during
DNA replication ultimately
results in the change of UGA
codon to UGG?
52
Post-translational Modifications
Phosphorylation
Nitrosylation Glycosylation
Post-
translational
modification
Methylation Proteolysis
Lipidation
53
Remember
When translating an mRNA sequence,
identify first the location of the start
codon AUG. From this, divide the
succeeding sequence into segments of
three nucleotides until you encounter
any of the three stop codons (UAA,
UAG, UGA).
54
Remember
For example, the given mRNA sequence
below can be divided correspondingly.
GUCAAUGGCGCCGUUUCGUUAGCGUCCACC
A
GUCA|1AUG(START)|2GCG|3CCG|4UUU|5CGU|
6
UAG(STOP)|CGUCCACCA
You can already disregard the rest of the
mRNA after the stop codon when looking for
the corresponding amino acids in the
55
Check Your
Understanding
Arrange the following steps in DNA replication
from the first process by labeling with numbers
from 1 to 10.
1.DNA polymerase I replaces the RNA primers with DNA
nucleotides.
2.In case of errors, DNA polymerase III performs a 3’ to 5’
exonuclease activity.
3.Primase synthesizes short RNA primers in both the leading
and the lagging strands.
56
Check Your
Understanding
Arrange the following steps in DNA replication
from the first process by labeling with numbers
from 1 to 10.
4.DNA polymerase III performs the majority of the DNA
nucleotide polymerization in both the leading and lagging
strands in 5’ to 3’ direction.
5.The initiator proteins unwind a short sequence at the
replication origin.
6.DNA helicase performs the rest of the unwinding of the DNA
helix by breaking hydrogen bonds.
57
Check Your
Understanding
Arrange the following steps in DNA replication
from the first process by labeling with numbers
from 1 to 10.
7.The initiator proteins recognize the location of the replication
origin.
8.The single-stranded binding proteins attach to and stabilize
the single strands of DNA.
9.DNA gyrase relieves the tension that builds up as the
helicase unwinds the DNA.
10.DNA ligase seals the nick between the nucleotides that are
synthesized by polymerase I and III.
58
Check Your
Understanding
In each of the following DNA sequences, write
the corresponding mRNA transcript and use the
genetic code to determine the resulting amino
acid sequence. You may proceed even without
the start codon.
1.TTTTACCATCCCACAATTTA
2.ACTACTTTCAGAGCTATATTCAG
3.CATTACGGAGCCTGATGCACTTAC
4.TACGCCGCAACTCCGTATGGC
5.GCTACAGCCCTAGCATTTACCCG
59
Let’s Sum It Up!
● The semiconservative DNA replication
involves the formation of a replication bubble
consisting of two replication forks on opposite
ends. New strands are synthesized in the 5’ to
3’ direction.
60
Let’s Sum It Up!
● At the replication origin, initiator proteins,
DNA helicase, DNA gyrase (or
topoisomerase II in humans), and single-
stranded binding proteins cooperate in
unwinding the DNA helix and maintaining
each of the strands available for replication.
61
Let’s Sum It Up!
● After the synthesis of primers, the DNA
polymerase III synthesizes new nucleotide
chains in the 5’ to 3’ direction. Synthesis is
continuous in the leading strand because
polymerase moves in the direction similar to
that of the unwinding of DNA. By contrast,
synthesis occurs in segments called Okazaki
fragments in the lagging strand because of
the antiparallel nature of the helix. 62
Let’s Sum It Up!
● DNA polymerase performs proofreading
mechanisms during replication. For errors that
are bypassed during replication, mismatch
repair is performed by proteins before further
proceeding to the G2 phase of the cell cycle.
63
Let’s Sum It Up!
● The central dogma of molecular biology
involves a series of processes that involve the
expression of genes into proteins. It serves as
the link between the genotype and phenotype.
64
Let’s Sum It Up!
● Transcription is the process that produces
RNA from the information in DNA. There are
different types of RNA molecules, but only
mRNA is translated into an amino acid
sequence. The other RNAs participate in either
the transcription and protein synthesis without
being further translated.
65
Let’s Sum It Up!
● The core enzyme of the RNA polymerase
then synthesizes the RNA nucleotide chain in
the 5’ to 3’ direction. Particularly, it only
transcribes the sense or template sequence of
the gene. The RNA polymerase also has the
ability to unwind and rewind the DNA as it
moves through the helix.
66
Let’s Sum It Up!
● The resulting mRNA molecule undergoes a
series of chemical modifications such as
methylation (5’ capping), polyadenylation
(addition of poly-A tail), and intron splicing.
67
Let’s Sum It Up!
● Translation is the synthesis of the
polypeptide by using the information in mRNA.
mRNA is read in triplets of nucleotides called
codons. The genetic code serves as the basis
of correspondence between the codons of
mRNA and the 20 essential amino acids.
68
Let’s Sum It Up!
● During translation, the large and small
ribosomal subunits, mRNA, and tRNAs
assemble to form the translation complex.
The tRNAs convey the individual amino acids
as they correspond to each of the codons. The
ribosomes recognize the mRNA sequence and
catalyze the elongation of the peptide chain.
69
Let’s Sum It Up!
Summary of the central
dogma of the cell
70
Challenge Yourself
If a codon will consist of four
nucleotides instead of three,
how will this affect the genetic
code?
71
Photo Credit
● Slide nos. 1 and 4: GFP Fluorescent Protein Movie by Erin Rod is licensed under
CC BY-SA 4.0 via Wikimedia Commons.
72
Bibliography
Alberts B., Bray D., Hopkin K., Johnson A., and Lewis J. Essential cell biology (4th ed). New
York, NY: Garland Science. 2014.
Klug, W.S, and Cummings, M.R. Concepts of genetics (6th ed). Upper Saddle River, N.J:
Prentice Hall. 2003.
Pierce, Benjamin. Genetics: a conceptual approach (8th ed). New York: W.H. Freeman. 2012.
Simmons, M.J. and Snustad, D.P. Principles of genetics (9th ed). Hoboken, NJ. Wiley. 2012.
Turner, P.C., McLennan, A.G., Bates, A.D., and White M.R.H. BIOS Instant Notes in Molecular
Biology (3rd ed). Garland Science, Taylor and Francis Group, LLC. 2005.
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