LECTURE 4
TRANSLATION……Continued
(PROTEIN SYNTHESIS)
Stages of Translation
• Translation which occurs in the cytoplasm of
eukaryotic cell is the second step in gene
expression which leads to protein synthesis.
• Protein synthesis via translation involves three
steps; initiation, elongation and termination.
1. Initiation
• Initiation is the process that brings all the
translation components together.
• These components include a proteins called
initiation factor required to assemble the
small and large ribosomal subunits, mRNA and
tRNA for the start of protein synthesis.
• In prokaryotes, the ribosome gets attached to
a strand of mRNA at the start codon AUG
which codes for the amino acid methionine.
• Although similar in many ways, initiation in
eukaryotes is much more complex and
complicated.
2. Elongation
• This is a step in which a polypeptide increases
in length by addition of one amino acid at a
time.
• It involves the participation of the transfer
RNA (tRNA)
• The tRNA anticondon recognises and decodes
an mRNA codon.
• The acceptor arm binds appropriate amino
acids on the 3’ carbon end of the molecule.
• The DHU arm bears enzyme complexes
(synthetases) that help with the attachment
of the amino acid at the 3’ carbon end.
• The T arm is the recognition site for the
ribosome to form the tRNA-ribosme complex.
• The order of the codons determines the order
of the tRNA molecules at a ribosome and the
sequence of amino acids.
• The AUG codon on mRNA gets lined up with
the peptidyl (P) site of the ribosome.
• The large ribosomal units consist of an amino
acyl site (A) where the incoming tRNA
carrying an amino acid attaches and peptidyl
site (P) where amino acids are joined through
peptide bonds and the exit site (E) where
tRNA leaves the ribosomal unit.
• Ribosomal(r) RNA plays a catalytic role in the
creation of a chemical bond (peptide bond)
between amino acid with the aid of an
enzyme called Peptidyl transferase .
• A transfer RNA (tRNA) with anticondon UAC
collects methionine to become the first amino
acid in the peptide chain.
• During elongation codons on mRNA are read
and amino acids are polymerised using
anticodons on tRNA as it move from A site to
P site to E site in a process called
translocation.
• mRNA moves forward into the ribosome
beginning with the 5’ end towards the 3’ end.
• mRNA is ready by tRNA in the 5’ to 3’
direction.
• Enzymes called aminoacyl-tRNA synthetases
catalyze the addition of amino acids to the
tRNAs.
• The tRNA with anticodon for Valine moves out
of the E site of the ribosome to fetch another
Valine from the cytoplasm.
• The tRNA with anticodon for lysine shifts to
the peptidyl position where codon AAA is
situated on the mRNA leaving the A site free.
• The tRNA with anticodon for Aspartic acid
occupies the free A site where the codon GAU
is located on the mRNA.
• The peptide continues to grow as it binds to
the incoming amino acid Tryptophan arriving
at site A until a stop codon UGG is read.
3. Termination
• Termination of a polypeptide synthesis occurs
when a specific nucleotide sequence is
encountered in the message on mRNA.
• This is when one of the three RNA triplet
codes- UAA, UAG or UGA are in the reading
of mRNA message.
• These triplets do not have complimentary
anticodons and therefore they do not have
an associated tRNA.
• They release a protein that help to dislodge
the complete polypeptide chain from the
ribosome.
• The ribosome dissociates into two subunits
and the separate from the mRNA.
• Soon after the mRNA is disintergrated by RNA
polymerase.
Post translational modification.
• Proteins are not yet complete and functional
when termination occurs.
• Most proteins go through an extensive
series of processing steps, collectively called
post-translational modification, before they
are completely functional.
• These modification include;
1. Structural modification (folding) :Although
folding can occur spontaneously, it is
frequently speeded up by proteins called
molecular chaperones.
2. Chemical modification: Chemical groups
may be added to proteins—often sugar or
lipid groups that are critical for normal
functioning of protein.
• This normally occurs in the golgi bodies and
endoplasmic reticulum.
• Adding or removing a phosphate group can
also cause major changes in the shape and
chemical reactivity of proteins.
• These changes have a dramatic effect on the
protein’s activity—often switching it from an
inactive state to an active state, or vice versa.
Gene Expression
• A gene is expressed once its product, a protein
is made and is functioning in a cell.
• For a protein, gene expression requires
transcription and translation.
• It also requires respective transcriptional
and translational modification.
• In addition completed and folded proteins
may be activated or deactivated by events
such as phosphorylation.
• The process of making protein is the same in
all organisms because they all use nucleic
acid.
• However the tRNA differs from one species to
another which is the basis for speciation.
• Error may occur during protein synthesis.
• This could be due to errors
i. During DNA replication
ii. Synthesis of mRNA
iii. Formation of complexes between the various
tRNA and amino acids and the interaction
between these complexes and the mRNA on
the ribosome.
END OF LECTURE!
THANK YOU.
