Lesson 2

Central Dogma: DNA Replication and

Protein Synthesis
SPECIFIC OBJECTIVES:
At the end of the lesson, the student is expected to:
• describe the requirements, proteins and enzymes in DNA replication;
• transcription and translation; and
• diagram the steps in replication, transcription and translation.

2.1 DNA Replication

Definition 2.1 DNA Replication

Before a cell divides, it copies its DNA so both descendant cells will inherit a
full complement of chromosomes. Replication of each DNA molecule produces two
duplicates. It is also semiconservative which means that the two “daughter” DNA
molecules will each be consists of one parental strand and one new strand.

Remarks:

2.1.1 Major Steps of Replication

a. Helicase: unwinds DNA at origins of
replication
b. Initiation proteins separate 2 strands
→ forms replication bubble

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 c. Primase: puts down RNA primer to
start replication

d. DNA polymerase III: adds

complimentary bases to leading
strand (new DNA is made 5’ > 3’)

e. Lagging strand grows in 3’→5’

direction by the addition of
Okazaki fragments
o Okazaki Fragments:
Short segments of DNA
that grow 5’→3’ that are
added onto the Lagging
Strand
f. DNA polymerase I: replaces
RNA primers with DNA
g. DNA ligase: seals fragments
together

2.1.2 Major proteins involved in replication and their function

Protein Function
Unwinds the DNA at the origin of
Helicase
replication creating replication forks
Binds to and stabilizes single stranded
Single-strand binding protein (SSB)
DNA until it can be used as a template
Synthesizes an RNA at 5’ end of leading
Primase strand and each Okazaki fragment of
lagging strand
Synthesizes new DNA strand by adding
DNA polymerase III nucleotides to the 3’ end of a pre-existing
DNA strand
Removes RNA nucleotides of primer from
DNA polymerase I 5’ end and replaces them with DNA
nucleotides
Joins 3’ end of DNA that replaces primer
DNA ligase to rest of leading strand and joins Okazaki
fragments of lagging strand
2.1.3 Mutations that occur during replication
Definition 2.1.3 Mutations

DNA damage by environmental agents can cause replication errors. Newly

forming DNA is monitored for errors, most of which are corrected. Uncorrected
errors become mutations.

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 a. Large scale mutations: chromosomal; always cause disorders
or death
• nondisjunction, translocation, inversions, duplications,
large deletions
b. Point mutations: alteration in 1 base pair of a gene (ex. Sickle-
cell Disease)
i. Base-pair substitutions – replace 1 with another
1. Missense: different amino acid
2. Nonsense: stop codon, not amino acid
ii. Frameshift – mRNA read incorrectly; nonfunctional
proteins
• Caused by insertions or deletions

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 2.1.4 Proofreading and Repair
• DNA polymerases proofread as bases added
o Mismatch repair: special enzymes fix incorrect
pairings
o Nucleotide excision repair:
-Nucleases cut damaged DNA
-DNA polymerase and ligase fill in gaps
-Errors:
• Pairing errors: 1 in 100,000 nucleotides
• Complete DNA: 1 in 10 billion nucleotides

2.2 Protein Synthesis

Definition 2.2 Protein Synthesis

One of the most fundamental biological processes by which individual cells

build their specific proteins. Gene Expression is the process by which DNA directs
the synthesis of proteins or RNAs.

Remarks:

The Genetic Code

• For each gene, one DNA strand is the
template strand (3’>5’)
• mRNA triplets (codons) code for amino
acids in a polypeptide chain
• The antisense strand of DNA is read by
RNA polymerase from the 3' end to the
5' end during transcription (3' → 5'). The
complementary RNA is created in the
opposite direction, in the 5' → 3'
direction, matching the sequence of the
sense strand with the exception of
switching uracil for thymine.

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 2.2.1 DNA to RNA: Transcription
Definition 2.2.1 Transcription

During transcription, a gene region in one strand of DNA serves as a

template for assembling a strand of RNA. In eukaryotes, a new RNA is modified
before leaving the nucleus. This process happens outside the nucleus/ in the
cytoplasm

A. Stages of Transcription
I. Initiation
a. Eukaryotes:
- TATA box – DNA sequence (TATAAAA) upstream
from promoter
- Transcription factors must recognize TATA box before
RNA polymerase can bind to DNA promoter
:

b. Prokaryotes: RNA polymerase binds directly to promoter

II. Elongation - RNA polymerase adds RNA nucleotides to the 3’ end of the
growing chain (A-U, G-C)
o As RNA polymerase moves, it untwists DNA, then rewinds it after
mRNA is made

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 III. Termination
RNA polymerase transcribes a terminator sequence in DNA, then mRNA
and polymerase detach.
a. Eukaryotes:
- It is now called pre-mRNA and will undergo RNA
processing
b. Prokaryotes = mRNA ready for use and will proceed directly to
Translation

B. RNA Processing
✓ Additions to pre-mRNA
o 5’ cap (modified Guanine) and 3’ poly-A tail are added.
It helps in exporting mRNA from nucleus, protect from enzyme
degradation and attach to ribosomes

✓ RNA Splicing
o Pre-mRNA has introns (noncoding sequences) and exons
(codes for amino acids)
o Splicing occurs when introns are cut out and exons are joined
together

o small nuclear ribonucleoproteins = snRNPs

▪ snRNPs join with other proteins to form a spliceosome
o Spliceosomes catalyze the process of removing
introns and joining exons

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 2.2.2 RNA to Protein: Translation

Definition 2.2.2 Translation

During translation, amino acids are assembled into a polypeptide in the order
determined by the sequence of codons in an mRNA. It occurs in the ribosomes.

A. Types of RNA involved in Translation

1. mRNA (Messenger RNA)
o Encodes amino acid sequence of a polypeptide.
2. tRNA (Transfer RNA)
o Brings amino acids to ribosomes during translation.
o Specific to each amino acid
o Anticodon: pairs with complementary mRNA
codon

3. rRNA (Ribosomal RNA)

o With ribosomal proteins, makes up the ribosomes,
the organelles that translate the mRNA.
o Ribosome = rRNA + proteins
o Consists of 2 subunits (small and large ribosomal
subunit)

B. Codon Table
o A codon is a sequence of three DNA or RNA nucleotides that corresponds
with a specific amino acid or stop signal during protein synthesis. DNA and
RNA molecules are written in a language of four nucleotides; meanwhile,
the language of proteins includes 20 amino acids.
o This code is universal: all life forms use the same code.
o 64 different codon combinations

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 C. Stages of Translation
I. Initiation
✓ The small ribosomal subunit binds to start codon (AUG) on mRNA. The
tRNA carrying Met attaches to P site then the large subunit will bind to
the small subunit forming the translation initiation complex.

II. Elongation
✓ The ribosome continues to translate each codon in turn. Each
corresponding amino acid is added to the growing chain and linked via
a bond called peptide bond.

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 III. Termination
✓ Occurs when the ribosome encounters a stop codon. At a stop codon, a
release factor reads the triplet, and polypeptide synthesis ends; the
polypeptide is released from the tRNA, the tRNA is released from the
ribosome, and the two ribosomal subunits separate from the mRNA.

Reference Book:
Biology: The Unity and Diversity of Life, 14th ed by Starr, Evers, & Starr (2014)

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