BIOCHEMIST

RY
22- Protein
Biosynthesis
PRESENTED BY:
DR. MOHAMMED SALEH
MBCHB
Dr. Mohammed
Saleh
MBChB
Baghdad University

Peaks Medical Academy

2
Protein Biosynthesis
Steps of Protein Synthesis
Protein Biosynthesis
DNA RNA PROTEIN
Central Dogma
■ Central Dogma: It
means that DNA
makes RNA and RNA
makes proteins
 Protein Synthesis
■ The two main processes involved in protein synthesis are
– the formation of mRNA from DNA (transcription)
– the conversion by tRNA to protein at the ribosome rRNA
(translation)
■ Transcription takes place in the nucleus, while translation takes
place in the cytoplasm
■ Genetic information is transcribed to form mRNA much the same
way it is replicated during cell division.
Transcription
■ The synthesis of RNA
requires:
– the presence of the
four nucleotide
triphosphate [ATP,
GTP, CTP and UTP].
– Templet.
– Enzyme (DNA
dependent RNA
polymerase enzyme).
 Transcription
 Several steps occur during transcription:
 A section of DNA containing the gene unwinds.
 One of the separated strands is used as a template for the synthesis of mRNA as a complementary
strand, copied starting at the initiation point (promotor), which has the sequence TATAAA.

 An mRNA is synthesized using

complementary base pairing with
uracil (U) replacing thymine (T), The
synthesis of mRNA is catalyzed by
DNA dependent RNA polymerase
enzyme.
 The newly formed mRNA moves out
of the nucleus to ribosomes in the
cytoplasm and the DNA re-winds
RNA Polymerase
■ During transcription, RNA polymerase moves along the DNA
template in the 3’-5’direction to synthesize the corresponding
mRNA is synthesized in 5’-3’direction.
■ The mRNA is released at the termination point
RNA Polymerase
■ There are three classes of RNA polymerase in the nucleus of
eukaryotic cells for the synthesis of RNA.
■ Each class of RNA polymerase recognizes particular types of
genes:
– RNA polymerase I: It synthesizes the precursor of the r-RNA
in the nucleolus.
– RNA polymerase II: It synthesizes the precursors of m-
RNA .
– RNA polymerase III: It produces the small RNA, including
tRNA and 5S rRNA.
Steps of RNA Synthesis (DNA
Transcription)
■ The process of transcribing a typical gene in an eukaryotic cell is divided
into 3 phases; initiation, elongation and termination.
1. Initiation: begins with binding of RNA polymerase to a region in the DNA
known as the promoter. In eukaryotic cells, each class of polymerases has
specific transcription factors and specific promotor areas. When a DNA
sequence in a specific gene is to be transcribed to mRNA using RNA
polymerase II, the ‘promoter’ consensus sequence is called TATA box.
2. Elongation: the promoter region has been recognized unwinding of DNA
helix occurs. RNA polymerase II begins to synthesize a transcript of the
DNA sequence. The elongation phase is said to begin when the transcript
exceeds 10 nucleotides in length.
3. Termination: The elongation of the single-stranded RNA chain continues
until a termination signal is reached. This leads to stop transcription.
Initiation
Elongation
Termination
Transcription Process
Post-transcriptional Processing
■ The mRNA formed and released from the DNA template is known as the
primary transcript.
■ In mammalian system, it undergoes extensive processing to become the
mature mRNA. These modifications such as:
– Poly-A Tailing at 3' End: This tail protects mRNA from attack by
exonuclease.
– Capping at 5' End: 'capped' at the 5' terminus by 7-methyl
guanosine triphosphate. The cap is useful in recognition of mRNA by
the translating machinery.
– Removal of introns.
– Splicing of exons (connect together). These processing occur mainly
in the nucleoplasm.
The Genetic code
■ It is clear from the way of synthesis of mRNA that the sequence
of bases in mRNA has been determined by the sequence of
bases in the gene in DNA.
■ Sequence of bases in the mRNA is known as codon.
■ This sequence determines the sequence of amino acids in the
protein.
■ Codons are presented in the messenger RNA (mRNA)
■ mRNA leaves the nucleus and transported to the ribosomes
carrying with it information in the form of base sequence.
Characteristics of the genetic code
1. Specificity: The genetic code is specific, that is a particular
codon always codes for the same amino acid.
2. Universality: The genetic code is universal, i.e the specificity
of the genetic code has been conserved from very early stages
of evolution.
3. Degeneracy: The genetic code is degenerate . Although each
codon corresponds to a single amino acid, a given amino acid
may have more than one triplet coding for it. For example,
arginine is specified by six different codons.
4. Nonoverlapping: The code is non-overlapping, the code is
read from a fixed starting point as a continuous sequence of
bases, taken three at a time i.e. a base which is read will not be
read again.
Characteristics of the genetic code
5. The code is triplet, i.e. composed of three bases. 61 of the 64
codon are associated with amino acids.
6. First codon ( AUG) codes for methionine is the initiation
codon of mRNA. ( AUG) = start codon .
7. Termination (“stop”) codons: Three of the codons [UAA;
UAG and UGA] act as stop codons. i.e. when protein
synthesis reaches one of these codons on mRNA, the
synthesis of protein is stopped.
Translation
■ Translation is the reading of coded
information in mRNA, and
converting it to a sequence of amino
acid in protein.
■ It is a translation, since it has
changed the sequence of base
language into that of amino acid
language.
■ The process of translation is started
by the coming of tRNA1-amino
acid1 carrying the first amino acid
to the ribosome.
Components Required for Translation
■ A large number of components are required for the synthesis of
a protein. These include:
1. Messenger RNA transcribed from DNA.
2. Transfer RNA to transport amino acids
3. Ribosomes RNA to read mRNA, align amino acids attached to
tRNA and great the peptide bonds between a.a.
4. Amino acids (20 a.a)
5. Protein factors and (enzymes) .
6. ATP and GTP are required as sources of energy.
Translation
■ Three bases in tRNA1 known
as anticodons form hydrogen
bonds with three bases in
mRNA known as codons.
■ Then comes tRNA2-amino
acid2 carrying the second
amino acid according to the
codon on mRNA.
■ tRNA2-aa becomes bound
with hydrogen bonds between
its anticodon bases and the
codon bases in mRNA.
Translation
■ An enzymatic reaction occurs between the carboxyl group of first amino
acid and the amino group of the second amino acid to form peptide bond.
The dipeptide remains joined to tRNA2, and tRNA1 leaves the site of
protein synthesis.
■ Then comes tRNA3-amino acid3, bringing the third amino acid
according to the codon on mRNA. A tripeptide is formed linked with
tRNA3, and tRNA2 leaves the site of protein synthesis.
■ The process continues by addition of amino acids according to the
sequence of codons on mRNA until protein synthesis is completed. The
completed polypeptide is then released from the last tRNA.
■ Protein synthesis starts with N-terminus and finished in the C-
terminus.
 Steps in Protein Synthesis
■ The mRNA is translated from its 5′-end to its 3′-end, producing a protein synthesized
from its amino-terminal end to its carboxyl-terminal end.
1. In eukaryotes, ribosomal subunit binds to the 5′-end cap of the mRNA and moves
down the mRNA until it encounters the initiator codon AUG. This “scanning” process
requires ATP.
2. The “Initiation codon” AUG is recognized by a special initiator tRNA. The initiator
tRNA enters the ribosomal site charged with methionine and goes directly to the site.
3. Elongation of the polypeptide chain involves the addition of a new a.a to the carboxyl
end of the growing chain. During elongation, the ribosome moves from the 5′-end to
the 3′-end of the mRNA that is being translated.
4. Termination It occurs when one of the three termination codons moves into the
ribosomal site.
Termination
■ After a polypeptide with all the amino ■ Therefore, protein synthesis
acids for a protein is synthesized, the ends (termination)
ribosome reaches the “stop” codon:
■ The polypeptide is released from
UGA, UAA, or UAG
the ribosome.
■ There is no tRNA with an anticodon
for the “stop” codons
THANK YOU
Dr. Mohammed Saleh