BIOKIMIA MOLEKULER

19 SEPTEMBER 2019
Translation (Protein Synthesis)
RNA  protein
Making a protein

• Many RNAs needed

• mRNA, tRNA,
rRNA
Translation
• Translation is the process of decoding the
mRNA into a polypeptide chain
• Ribosomes read mRNA three bases or 1
codon at a time and construct the proteins

4
Transcription

Translation

5
Step 1- Initiation
• mRNA transcript start
codon AUG attaches to
the small ribosomal
subunit
• Small subunit attaches to
large ribosomal subunit

mRNA transcript
8
Ribosomes

Large
subunit
P A
Site Site

mRNA

A U G C U A C U U C G
Small
subunit 9
 Step 2 - Elongation
• As ribosome moves, two tRNA with
their amino acids move into site A and
P of the ribosome
• Peptide bonds join the amino acids

10
 Initiation
aa2
aa1

2-tRNA
1-tRNA
G A U
anticodon U A C
hydrogen A U G C U A C U U C G A
bonds codon mRNA
copyright cmassengale 11
 Elongation
peptide bond
aa3
aa1 aa2

3-tRNA

1-tRNA 2-tRNA G A A
anticodon U A C G A U
hydrogen A U G C U A C U U C G A
bonds codon mRNA
12
 aa1 peptide bond
aa3
aa2

1-tRNA

U A C 3-tRNA
(leaves)
2-tRNA G A A

G A U
A U G C U A C U U C G A
mRNA

Ribosomes move over one codon 13

 peptide bonds
aa1 aa4

aa2 aa3

4-tRNA

2-tRNA 3-tRNA G C U

G A U G A A
A U G C U A C U U C G A A C U
mRNA

14
 peptide bonds
aa1 aa4
aa2

aa3

2-tRNA
4-tRNA
G A U
(leaves) 3-tRNA G C U

G A A
A U G C U A C U U C G A A C U
mRNA

Ribosomes move over one codon 15

 peptide bonds aa5
aa1
aa2
aa4
aa3

5-tRNA

U G A
3-tRNA 4-tRNA

G A A G C U
G C U A C U U C G A A C U
mRNA

16
 aa1 peptide bonds aa5
aa2
aa3
aa4

5-tRNA

3-tRNA U G A
G A A 4-tRNA

G C U
G C U A C U U C G A A C U
mRNA

Ribosomes move over one codon 17

 aa5
aa4 aa199 Termination
aa3 primary aa200
structure
aa2 of a protein

aa1
terminator
200-tRNA or stop
codon

A C U C A U G U U U A G
mRNA

18
End Product –The Protein!
• The end products of protein synthesis
is a primary structure of a protein
• A sequence of amino acid bonded
together by peptide bonds

aa5
aa3 aa4
aa2 aa199

aa1 aa200
19
Messenger RNA (mRNA)
start
codon

mRNA A U G G G C U C C A U C G G C G C A U A A

codon 1 codon 2 codon 3 codon 4 codon 5 codon 6 codon 7

protein methionine glycine serine isoleucine glycine alanine stop

codon

Primary structure of a protein

aa1 aa2 aa3 aa4 aa5 aa6

peptide bonds 20
Genetic Code
• Proteins are composed of 20 different amino acids
• A sequence of 3 nucleotides is used to code each amino
acid
• Each triplet of nucleotides is called a codon
• Start codon AUG codes for amino acid methionine
• 3 stop codons (UAG, UAA, UGA)
• There are 64 codons in the genetic code 43=64
• Several different codons can code for the same aa, but no
codon ever has more than one amino acid counterpart.
• Codons are always written in the form of the RNA
transcript from the original DNA molecule.
 Do we need new mRNA every time?

• NO
• The same mRNA may be used to
make many copies of the same
protein.
Termination of Protein Synthesis

• Translocation of the ribosome exposes a stop

codon in the A site. Stop codons do not code for
an amino acid, there are no corresponding tRNAs.
• A protein called a release factor binds to the
exposed A site causing the polypeptide to
separate from the remaining tRNA molecule
• Ribosome falls of the mRNA and translation stops
Termination of Protein Synthesis
Translation Details
• Translation begins when the mRNA codon “AUG” is
read by a ribosome
• Ribosome reads one codon at a time
AUG = methionine
GCU = alanine
• tRNA carries over the proper amino acid
• tRNA anticodon matches with the mRNA codon
• Prevents delivery of wrong amino acid
• One by one, amino acids are linked together
• Translation ends when a “stop” codon is reached
• What just happened?: A ribosome made a protein
 Now the codon and
anticodon match. This
ensures
Thisthe proper
is why the
amino acid (serine)
anticodon is is
delivered.
important!

Because the codon and anticodon don’t match, the wrong amino acid will
not be delivered.
Questions to answer:
1) In order, list the amino acids that will be delivered to this ribosome.

2) What is the anticodon of each codon?

tRNA tRNA tRNA tRNA tRNA tRNA

UAC GGC UCA GGU CCU AGA
3) When finished, how many amino acids in size is this protein?

Methionine Proline Serine Proline Glycine Serine

(MET) (PRO) (SER) (PRO) (GLY) (SER)

tRNA tRNA tRNA tRNA tRNA tRNA

UAC GGC UCA GGU CCU AGA
ignore
GAU AUG CCG AGU CCA GGA UCU UGA
start stop
Ribosome
Practice Problem
Always use mRNA to find
the amino acid

DNA mRNA tRNA Amino Acid

codon anticodon
Practice Problem
Always use mRNA to find
the amino acid

DNA mRNA tRNA Amino Acid

codon anticodon
Practice Problem
Always use mRNA to find
the amino acid

DNA mRNA tRNA Amino Acid

codon anticodon

C G C
C G C
A U A
mRNA codes
Translation: Initiation
• mRNA binds to a ribosome, and the transfer RNA corresponding to
the START codon binds to this complex. Ribosomes are composed of
2 subunits (large and small), which come together when the
messenger RNA attaches during the initiation process.
Translation: Elongation
• Elongation: the ribosome moves down the messenger RNA, adding new amino acids to the
growing polypeptide chain.
• The ribosome has 2 sites for binding transfer RNA. The first RNA with its attached amino
acid binds to the first site, and then the transfer RNA corresponding to the second codon
bind to the second site.
Translation: Elongation
• The ribosome then removes the amino acid from the first transfer RNA and
attaches it to the second amino acid.
• At this point, the first transfer RNA is empty: no attached amino acid, and the
second transfer RNA has a chain of 2 amino acids attached to it.
Translation: Termination
• The elongation cycle repeats as the ribosome moves down the messenger
RNA, translating it one codon and one amino acid at a time.
• The process repeats until a STOP codon is reached.
Polyribosomes
• A number of ribosomes can translate a single mRNA
molecule simultaneously forming a polyribosome
• Polyribosomes enable a cell to make many copies of a
polypeptide very quickly Completed
Growing polypeptide
polypeptides
Incoming
ribosomal
subunits
Start of End of
mRNA mRNA
(5 end) (3 end)
(a) An mRNA molecule is generally translated simultaneously
by several ribosomes in clusters called polyribosomes.

Ribosomes
mRNA

0.1 µm
This micrograph shows a large polyribosome in a prokaryotic
cell (TEM).
 Polyribosomes:
several ribosomes can simultaneously translate
the same mRNA molecule
 Prokaryotes vs. Eukaryotes
Prokaryotes:
1. Translation takes place directly after transcription
2. mRNA is not modified
3. Transcription and translation take place in the same area
 Prokaryotes vs. Eukaryotes
Eukaryotes:
1. Transcript is modified before leaving the nucleus (5’ cap and 3’
poly-A tail)
2. Modifications increase translation efficiency and lifespan of the
mRNA
3. Translation takes place on ribosomes located in the rough
endoplasmic reticulum (translation is physically separated from
transcription)
 Post-translation
• The new polypeptide is now floating loose in the cytoplasm if translated by a
free ribosome.
• Polypeptides fold spontaneously into their active configuration, and they
spontaneously join with other polypeptides to form the final proteins.
• Often translation is not sufficient to make a functional protein, polypeptide
chains are modified after translation
• Sometimes other molecules are also attached to the polypeptides: sugars,
lipids, phosphates, etc. All of these have special purposes for protein function.
Gene Expression is
controlled at all of these
steps:
•DNA packaging
•Transcription
•RNA processing and
transport
•RNA degradation
•Translation
•Post-translational

Fig 16.1
Some proteins function in the cytoplasm; others need to be
transported to various organelles.
How can proteins be delivered to
their appropriate destinations?
Proteins are directed
to their destinations
via signals in the
amino acid sequence
Protein Destinations: secretion or membrane
• Signal sequences target proteins for secretion
Translation of secreted proteins
Translation of membrane bound proteins
Translation of secreted or membrane
bound proteins

This step determines secretion or

membrane bound.
Protein Destinations: nucleus Signal anywhere in protein,
Translation in cytoplasm,
Signal not removed
 Protein Destinations: mitochondria or
chloroplast

Signal translated first, Translation in cytoplasm, Signal removed

Protein Destinations: signals in protein
determine destination