DNA RNA
TRANSCRIPTION
Chapter 12: From DNA to Protein: Genotype to Phenotype
TOPICS
The central dogma
DNA structure
DNA replication
RNA structure
RNA synthesis (Transcription)
The genetic code
Protein synthesis (Translation)
Chapter 12: From DNA to Protein: Genotype to Phenotype
DNA and RNA differ
RNA differs from DNA in three ways:
RNA is single-stranded (but it can
fold back upon itself to form
secondary structure, e.g. tRNA)
In RNA, the sugar molecule is
ribose rather than deoxyribose
In RNA, the fourth base is uracil
rather than thymine.
Chapter 12: From DNA to Protein: Genotype to Phenotype
DN RN
A A
U
3
OH
OH 2
OH
OH
1
Chapter 12: From DNA to Protein: Genotype to Phenotype
The Central Dogma
The Flow of Information: DNA RNA
protei
n DNA Replication
Transcription Translation
RNA is synthesized via a process called
Transcription
mRNA, rRNA and tRNA are transcribed by
similar mechanisms
Three types of RNA are involved in
Chapter 12: From DNA to Protein: Genotype to Phenotype
protein synthesis
1. Messenger RNA
[mRNA]
- the template
2. Ribosomal RNA
[rRNA]
- structural component of
the ribosome
3. Transfer RNA [tRNA]
- the adapter
Chapter 12: From DNA to Protein: Genotype to Phenotype
Chapter 12: From DNA to Protein: Genotype to Phenotype
TRANSCRIPTION: DNA-Directed
RNA Synthesis (@ NUCLEUS)
Initiation
Elongation
Termination
RNA is transcribed from a DNA template after
the bases of DNA are exposed by unwinding
of the double helix.
In a given region of DNA, only one of the
two strands can act as a template for
transcription.
DURING DNA REPLICATION…,
Chapter 12: From DNA to Protein: Genotype to Phenotype
Transcription: DNA-Directed RNA
Synthesis - Initiation
INITIATION: RNA
polymerase recognizes and binds to
a promoter sequence on DNA.
The PRIBNOW BOX SEQUENCE is a
sequence of TATAAT of six nucleotides
that is an essential part of a promoter
site on DNA for transcription to occur in
bacteria.
PRIBNOW BOX SEQUENCE
Chapter 12: From DNA to Protein: Genotype to Phenotype
Transcription: DNA-Directed RNA
Synthesis - Elongation
ELONGATION: RNA polymerase
elongates the nascent RNA molecule in a 5’-
to-3’ direction, antiparallel to the template
DNA
• Nucleotides are added by
complementary base pairing with the
•
template strand.
The substrates, ribonucleoside
triphosphates, are hydrolyzed as added,
releasing energy for RNA synthesis.
Chapter 12: From DNA to Protein: Genotype to Phenotype
Transcription: DNA-Directed RNA
Synthesis - Termination
TERMINATION: Special DNA
sequences and protein helpers terminate
transcription.
The transcript is released from the DNA.
This Primary Transcript is called the “pre-
mRNA”
The pre-mRNA is processed to generate the
mature mRNA
TERMINATION (prokaryotes)
The Ras homologous (Rho) protein family is a member
of the Ras superfamily of small GTPases. Small GTPases
are monomeric proteins and function as molecular
switches that turn “on” or turn “off” signal transduction
pathways in response to chemical or mechanical stimuli
Termination of RNA Transcript can be:
RHO INDEPENDENT
RHO DEPENDENT
RHO INDEPENDENT
TERMINATION
RHO DEPENDENT TERMINATION
RHO DEPENDENT TERMINATION
Chapter 12: From DNA to Protein: Genotype to Phenotype
Figure 12.4 – Part 2
Chapter 12: From DNA to Protein: Genotype to Phenotype
Transcription: DNA-Directed RNA
Synthesis - Termination
RNA PROCESSING
From Pre mRNA to
Mature mRNA
Chapter 12: From DNA to Protein: Genotype to Phenotype
Transcription: DNA-Directed RNA
Synthesis – RNA PROCESSING
Transcribed RNA is known as the pre-
mRNA. It is processed further to
convert it into mature RNA. RNA
processing include:
Capping
Polyadenylation
Splicing
Chapter 12: From DNA to Protein: Genotype to Phenotype
Transcription: DNA-Directed RNA
Synthesis – RNA PROCESSING
Capping
A methylated guanine cap is added to
protect the mRNA. It involves:
Addition of methylated guanine
It occurs at 5′ end of mRNA transcript
It protects the mRNA from
degradation.
Chapter 12: From DNA to Protein: Genotype to Phenotype
Transcription: DNA-Directed RNA
Synthesis – RNA PROCESSING
Capping
Chapter 12: From DNA to Protein: Genotype to Phenotype
Transcription: DNA-Directed RNA
Synthesis – RNA PROCESSING
Polyadenylation
The poly-A tail also protects the
mRNA from degradation. It involves:
The endonucleases cleave the mRNA
at a specific sequence.
The enzyme polyA polymerase
facilitates the addition of several
adenine nucleotides.
POLYADENYLATION of
mRNA
POLYADENYLATION of
mRNA
Chapter 12: From DNA to Protein: Genotype to Phenotype
Transcription: DNA-Directed RNA
Synthesis – RNA PROCESSING
Splicing
The non-coding sequences, i.e., the
introns are removed by spliceosome
excision.
The coding sequences or the exons
join together by ligation.
snRNP’s
Chapter 12: From DNA to Protein: Genotype to Phenotype
Topics
The central dogma
DNA structure
DNA replication
RNA structure
RNA synthesis (Transcription)
The genetic code
Protein synthesis (Translation)
RNA PROTEIN
TRANSLATION
Chapter 12: From DNA to Protein: Genotype to Phenotype
The Central Dogma
The Flow of Information: DNA RNA
protei
n DNA Replication
Transcription Translation
A gene is expressed in two steps:
DNA is transcribed to RNA
Then RNA is translated into protein.
Genetic Code
Genetic code contains the information of the
protein manufactured from RNA.
TRIPLET CODONS: t nucleotides form a triplet
codon that codes for one amino acid.
The number of possible amino acids range to 4 x 4
x 4 = 64 amino acids. There are 20 naturally
existing amino acids.
Chapter 12: From DNA to Protein: Genotype to Phenotype
TRANSLATION
Translation- the synthesis of protein
from an RNA template.
(@CYTPLASM)
Five stages: Pre-initiation
1. Initiation
2. Elongation
3. Termination
Post-translational modification
Complicated: In eukaryotes, ~300
molecules involved
Chapter 12: From DNA to Protein: Genotype to Phenotype
FUNCTIONS OF THE TYPES OF
RNA
mRNA- serves as a template code
tRNA- serves as an adapter
molecule
rRNA- holds molecules in the
correct
position, protein portion also
catalyze reactions
Chapter 12: From DNA to Protein: Genotype to Phenotype
tRNA Structure
All tRNA molecules have a similar but not identical
structure- “cloverleaf”
Chapter 12: From DNA to Protein: Genotype to Phenotype
tRNA
Structure
Chapter 12: From DNA to Protein: Genotype to Phenotype
PRE-INITIATION - Charging
the tRNA
1. Charging the tRNA
2. Formylation of met-tRNAfmet
Charging the tRNA
Charging the tRNA
Chapter 12: From DNA to Protein: Genotype to Phenotype
PRE-INITIATION - Charging
the tRNA
1. Formylation of met-tRNAfmet
TETRAHYDROFOLATE
Chapter 12: From DNA to Protein: Genotype to Phenotype
rRNA
RIBOSOME COMPOSED OF 2 SUBUNITS:
Small subunit – matches the tRNAs to the codons of
the mRNA
Large subunit – catalyzes the formation of the peptide
bonds between amino acids in the growing
polypeptide chain
The two subunits come together near the 5’ end of
the mRNA to begin synthesis of a protein
Then ribosome moves along, translating codons, until
2 subunits separate after finishing.
Chapter 12: From DNA to Protein: Genotype to Phenotype
Each ribosome contains 3
binding sites for tRNA
molecules:
A-site = aminoacyl-tRNA
P-site = peptidyl-tRNA
E-site = exit
SITES OF TRANSLATION
Chapter 12: From DNA to Protein: Genotype to Phenotype
Mechanism for finding start codon is
different in bacteria
Instead of a 5’ cap, mRNA has specific
ribosome- binding sequence located
upstream of AUG = Shine-
Dalgarno sequence
Chapter 12: From DNA to Protein: Genotype to Phenotype
INITIATION: mRNA Structure
Shine-Dalgarno sequence
~10 nt upstream of initiation codon
- Positions ribosome at correct start
site
Chapter 12: From DNA to Protein: Genotype to Phenotype
The initiator tRNA is loaded into the P site
of ribosome along with translation
initiation factors
The loaded ribosomal small subunit binds
to the 5’ end of the mRNA, recognized by
the cap
Then moves forward along the mRNA
searching for the AUG
Once found, large subunit associates
Protein synthesis begins with next tRNA
binding to the A site, etc.
Chapter 12: From DNA to Protein: Genotype to Phenotype
Codons in mRNA signal where to start
and stop protein synthesis
Translation begins with codon AUG (start
codon) and a special tRNA required for
initiation—
The initiator tRNA always carries
methionine (Met) or a modified form
of it
START CODON : AUG
START ANTICODON : UAC
INITIATION
Step 2
Step 1
INITIATION
All INITIATION FACTOR (IF)
are released
ELONGATION
AMINO
ACID
E P A
Chapter 12: From DNA to Protein: Genotype to Phenotype
Termination
Chapter 12: From DNA to Protein: Genotype to Phenotype
RELEASE FACTOR
A protein release factor, rather than a
tRNA, binds to a stop codon
This signals peptidyl transferase to
add water rather than an amino acid
to the end of the growing polypeptide
Chapter 12: From DNA to Protein: Genotype to Phenotype
RELEASE FACTOR
07_34_stop codon.jpg
Chapter 12: From DNA to Protein: Genotype to Phenotype
RELEASE FACTOR
This releases that last amino acid
from the tRNA, and thus the
polypeptide from the ribosome
The ribosome releases the
mRNA and disassociates into
its 2 subunits
ANOTHER….
