Protein synthesis

(Translation of mRNA)

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Teaching aims
By the end of the lecture:
students are expected to understand
the molecular mechanism of translation
or protein synthesis
students are expected to understand
the regulation of gene expression

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Core topics

Overview
Translation of mRNA
Mutation
Regulation of gene expression

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Transfer of genetic information

Replication of DNA
transmission of genetic information
from parental cell to its daughter cells
Transcription of DNA
transmission of genetic information
from DNA to RNA
Translation of RNA (polypeptide/protein
biosynthesis)
transmission of genetic information
from RNA to polypeptide/protein
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Human Genome Size

NUCLEAR GENOME
* 23 pairs of chromosomes 2
x ( 3 x 109 b.p) 2 meters
DNA / Cell
* 2 x ( 3 x 1012 cells)
meters DNA in human body
8,000 x earth to moon
MITOCHONDRIAL GENOME
* circular, 16,569 bp
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One of the most

important stages in
RNA processing is
RNA splicing. In
many genes, the
DNA sequence
coding for proteins,
or "exons", may be
interrupted by
stretches of noncoding DNA, called
"introns".
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Translation of RNA
The ribosome binds to the mRNA at the
start codon (AUG) that is recognized
only by the initiator tRNA. The ribosome
proceeds to the elongation phase of
protein synthesis. During this stage,
complexes, composed of an amino acid
linked to tRNA, sequentially bind to the
appropriate codon in mRNA by forming
complementary base pairs with the tRNA
anticodon.
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 The ribosome moves from codon to

codon along the mRNA. Amino acids are
added one by one, translated into
polypeptidic sequences dictated by DNA
and represented by mRNA. At the end, a
release factor binds to the stop codon,
terminating translation and releasing the
complete polypeptide from the ribosome.

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Codon

Three-letter code words ( a triplet code)

Unambiguous
Non-overlapping
Without punctuation
Universal
Can be found either in DNA (sense
strand) and mRNA

The collection of codons (64) makes up the genetic code

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Three
nonsense
codons (UAA,
UAG, UGA) do
not code for
specific amino
acid and are
utilized as
termination
signal

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A = adenine G = guanine C = cytosine

T = thymine U = uracil
DNA transfers information to mRNA in the
form of a code defined by a sequence of
nucleotides bases.
During protein synthesis, ribosomes move
along the mRNA molecule and "read" its
sequence three nucleotides at a time
(codon) from the 5' end to the 3' end.
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 Each amino acid is specified by the

mRNA's codon, and then pairs with a
sequence of three complementary
nucleotides carried by a particular tRNA
(anticodon).
Since RNA is constructed from four
types of nucleotides, there are 64
possible triplet sequences or codons
(4x4x4).
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 Three of these possible codons specify

the termination of the polypeptide
chain. They are called "stop codons
(nonsense codons). That leaves 61 codons
to specify only 20 different amino acids.
Therefore, most of the amino acids are
represented by more than one codon.
The genetic code is said to be
degenerate.

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Amino acids specified by each codon sequence on mRNA

Ala: Alanine

Cys:
Cysteine

Asp: Aspartic Glu: Glutamic

acid
acid

Phe:
Phenylalanine Gly: Glycine

Ile:
His: Histidine
Isoleucine

Lys: Lysine

Met:
Leu: Leucine
Methionine

Asn:
Asparagine

Pro: Proline

Gln:
Glutamine

Arg: Arginine

Ser: Serine

Thr:
Threonine

Val: Valine

Trp:
Tryptophane
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Tyr: Tyrosisne

Protein translation takes place by the

following steps
1. Formation of the initiation complex
2. Elongation of the polypeptide chain (one
repetition of the steps a, b and c for every
amino acid incorporated into the protein being
made):
a. binding of aminoacyl-tRNA
b. peptide bond formation
c. translocation
3. Termination
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Remember !
Proteins are polypeptides made up of
individual amino acids linked together,
Carbohydrates are polysaccharides
made up of individual monosaccharides
linked together, and
Nucleic acids are polynucleotides made
up of individual nucleotides linked
together.
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Mutations
Result when changes occur in the
nucleotide sequence may not occur in
the template strand but appear after
replication
Some mutations occur by base substitution
single base changes (point mutations):
Transitions (pryrimidine to other
pyrimidine, purine to other purine)
Transversion (pyrimidine to purine or
purine to pyrimidine)
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 Single base changes in DNA sequence

followed by changes in mRNA molecules may
have one of several effects when
translated into protein:
No detectable effect silent mutation
Missense effect missense mutation
Appearance of nonsense codon that result
in premature termination of polypeptide
chain being synthesized nonsense
mutation
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 Substitution of amino acids in protein

causes missense mutations (illustration on
Hemoglobin molecule):
Acceptable missense mutations
Hb Hikari: AAA or AAG (lys) to AAU or AAC
(asp)
Hb E: GAA or GAG (glu) to AAA or AAG (lys)
Partially acceptable missense mutations
Hb S: GAA or GAG (glu) to GUA or GUG (val)
Unacceptable missense mutations
Hb M: Hb (Fe2+) to met Hb (Fe3+)
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 Frameshift mutations result from

deletion or insertion of nucleotides
generates altered mRNAs
May be one, two, three or multiples
nucleotides

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Regulation of gene expression

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 In bacteria & viruses:

Alteration of gene expression is
required by organism to adapt to
environmental changes involves
interaction of specific binding proteins
with various regions of DNA in the
immediate vicinity of the transcription
start site

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 In eukaryotes:
in addition to those proteins,
alteration of gene expression also
involves tissue specific expression;
regulation by hormones, metals &
chemicals; gene amplification; gene
rearrangement; posttranscriptional
modification

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Type of responses to a regulatory signal

Type A: increased rate of gene
expression that is dependent upon the
continued presence of inducing signal
Type B: increased rate of gene
expression that transient even in the
continued present of regulatory signal
Type C: increased rate of gene
expression that persist indefinitely even
after the termination of the signal
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Type of gene regulation

Positive regulation:
The expression of genetic info is
quantitatively increased by the presence
of a specific regulatory element (the
molecule is positive regulator)
Negative regulation:
the expression of genetic info is
diminished by the presence of a specific
regulatory element (the molecule is
negative regulator)
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Model of gene expression in prokaryotes

One cistron, one subunit concept instead
of one gene one enzyme concept (cistron
is the smallest unit of gene expression,
coding for the structure of the subunit
of a protein molecule)
Inducible genes: their expression
increases in response to an inducer
Constitutive genes: their expression is
reasonably constant (not known to be
subject to regulation)
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The earliest level of regulation is at DNA level during transcription

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