Gene Expression - Protein Synthesis
Gene Expression - Protein Synthesis
Gene Expression - Protein Synthesis
Lecture objectives
• Describe how information encoded in genes
is used in the synthesis of proteins, through
– Transcription
– Translation
Central dogma of molecular biology
Protein Synthesis
• The information contained by DNA is in the form of
specific sequences of nucleotides along the DNA
strands (Genes)
• DNA in cells dictates synthesis of proteins
• The process by which DNA directs protein synthesis
(gene expression) includes two stages, called
transcription and translation
Protein Synthesis
Transcription and Translation
• In a eukaryotic cell the nuclear envelope separates transcription
from translation
• Extensive mRNA processing occurs in the nucleus
Nuclear
envelope
TRANSCRIPTION DNA
Pre-mRNA
RNA PROCESSING
mRNA
Ribosome
TRANSLATION
Polypeptide
Transcription and Translation
• Learning Outcomes
1. Describe the transcription process in
bacteria.
2. Differentiate features of initiation from those
of elongation.
3. Define the unique features of prokaryotic
transcription
Prokaryotic Transcription
• Prokaryotes have a single RNA
polymerase which exists in two
forms: the core polymerase and
the holoenzyme.
• core polymerase
• composed of four subunits: two
identical α subunits, a β
subunit, and a β' subunit.
• synthesizes RNA using a DNA
template, The active site of the
enzyme is formed by the β and
β' subunits, which bind to the
DNA template.
• holoenzyme
• is formed by the addition of a σ
(sigma) subunit to the core
polymerase. Has the ability to
recognize specific signals in
DNA allowing RNA polymerase
to locate the beginning of
genes.
• can accurately initiate
synthesis.
Prokaryotic Transcription:
Synthesis of an RNA Transcript
• The stages of Promoter
Transcription unit
transcription are 5 3
3 5
– Initiation Start point
DNA
RNA synthesis. DNA transcript 2 Elongation. The polymerase moves downstream, unwinding the
DNA and elongating the RNA transcript 5 → 3 . In the wake of
transcription, the DNA strands re-form a double helix.
Rewound
RNA
5 3
3 3 5
5
RNA
transcript
3 Termination. Eventually, the RNA
transcript is released, and the
polymerase detaches from the DNA.
5 3
3 5
5 3
Completed RNA
transcript
Prokaryotic Transcription:
Synthesis of an RNA Transcript- Termination
• Translation involves
TRANSLATION
Polypeptide
– mRNA
Amino
– Ribosomes - Ribosomal RNA acids
Polypeptide
– Transfer RNA
– Codons tRNA with
amino acid
Ribosome attached
synthesis 5 Codons 3
mRNA
Translation
• Before amino acids join together TRANSCRIPTION DNA
Gly
tRNA
Anticodon
A A A
U G G U U U G G C
5 Codons 3
mRNA
Prokaryotic Translation
Transfer RNA
• Consists of a single RNA strand that is only about 80 nucleotides long
• Each carries a specific amino acid on one end and has an anticodon on
the other end
• A special group of enzymes pairs up the proper tRNA molecules with
their corresponding amino acids.
• tRNA brings the amino acids to the ribosomes,
3
Amino acid A
C
attachment site C
The “anticodon” is the 3 RNA bases that A
C G
5
matches the 3 bases of the codon on the G
C
C
G
mRNA molecule U G
U A
A U
U C A U
* C A C AG UA AG *
G * CUC *
C GU GU* CGAG G
* * U C * AG G
* GAG C Hydrogen
G C
U A bonds
* GA
A* C
*A U
A G
Anticodon
Prokaryotic Translation
Transfer RNA and the charging
reaction
• Each amino acid must be attached to a
tRNA with the correct anticodon for protein
synthesis to proceed.
• This covalent attachment is accomplished
by the action of activating enzymes called
aminoacyl-tRNA synthetases.
– Cells produce 20 different aminoacyl-tRNA
synthetase enzymes, 1 for each of the 20 distinct
amino acids.
• Aminoacyl-tRNA synthetases recognizes a
specific amino acid and also ATP then
correct tRNA binds to the synthetase.
• the tRNA with its attached amino acid is
released from the enzyme. At this stage, the
tRNA is called a charged tRNA or an
aminoacyl-tRNA.
Prokaryotic Translation
• named ribosomal RNA
Ribosomes
Building a Polypeptide
Amino end Growing polypeptide
tRNA
mRNA 3
Codons
5
(c) Schematic model with mRNA and tRNA. A tRNA fits into a binding site when its anticodon base-
pairs with an mRNA codon. The P site holds the tRNA attached to the growing polypeptide. The A
site holds the tRNA carrying the next amino acid to be added to the polypeptide chain. Discharged
tRNA leaves via the E site.
Prokaryotic Translation
Building a Polypeptide
Initiator tRNA
GTP GDP
E A
mRNA
5 3 5 3
Start codon
1 2
A small ribosomal subunit binds to a molecule of The arrival of a large ribosomal subunit completes
mRNA. In a prokaryotic cell, the mRNA binding site the initiation complex. Proteins called initiation
on this subunit recognizes a specific nucleotide factors (not shown) are required to bring all the
sequence on the mRNA just upstream of the start translation components together. GTP provides
codon. An initiator tRNA, with the anticodon UAC, the energy for the assembly. The initiator tRNA is
base-pairs with the start codon, AUG. This tRNA in the P site; the A site is available to the tRNA
carries the amino acid methionine (Met). bearing the next amino acid.
•
Prokaryotic Translation
In the elongation stage, amino acids are added one by one to
the preceding amino acid
Elongation of the Polypeptide Chain
1 Codon recognition. The anticodon
TRANSCRIPTION DNA
Amino end of an incoming aminoacyl tRNA
mRNA
of polypeptide base-pairs with the complementary
Ribosome
TRANSLATION mRNA codon in the A site. Hydrolysis
Polypeptide
of GTP increases the accuracy and
E efficiency of this step.
mRNA 3
Ribosome ready for P A
next aminoacyl tRNA 5 site site
2 GTP
2 GDP
E E
P A P A
Release
factor
Free
polypeptide
5
3 3
3
5 5
Stop codon
(UAG, UAA, or UGA)
1 When a ribosome reaches a stop 2 The release factor hydrolyzes 3 The two ribosomal subunits
codon on mRNA, the A site of the the bond between the tRNA in and the other components of
ribosome accepts a protein called the P site and the last amino the assembly dissociate.
a release factor instead of tRNA. acid of the polypeptide chain.
The polypeptide is thus freed
from the ribosome.
Eukaryotic Transcription
Learning Outcomes
• List the different eukaryotic RNA polymerases.
• Distinguish between the promoters of the RNA
polymerases.
• Define the processing that occurs to eukaryotic
transcripts.
Eukaryotic Transcription
3
RNA
5 RNA polymerase
transcript
RNA PROCESSING
Exon
Summary of RNA transcript
transcription (pre-mRNA)
in a eukaryotic Aminoacyl-tRNA
synthetase
cell NUCLEUS
Amino
acid AMINO ACID ACTIVATION
CYTOPLASM tRNA
mRNA Growing
polypeptide
3
A
Activated
P amino acid
Ribosomal
E
subunits
5
TRANSLATION
E A
Anticodon
Codon
Ribosome