Translation in Prokaryotes

The genetic material of prokaryotes is not contained in a membrane-bound nucleus,

transcription takes place in the cytoplasm. This, then, allows translation to begin in
this environment as soon as mRNA emerges from the polymerase.

As a result, a scenario in which a strand of DNA is being transcribed by multiple

polymerases with multiple ribosomes translating this information of the RNA.
Initiation

For the initiation phase to take place, the smaller ribosomal subunit first has to be
dissociated from the larger ribosomal subunit. Once it's dissociated, initiation factors
bind at given sites on the 30S subunits where they serve different functions.

At the A site (of the ribosome subunit), initiation factor serves to prevent a new
molecule of tRNA from entering at this stage of translation. In addition, it promotes the
assembly and stabilization of the complex.

As well, the initiation factor promotes binding of the subunit to mRNA. The third
initiation factor introduces the initiator aminoacyl-tRNA and binds the P site of the
subunit. In doing so, it allows the anticodon of the tRNA to attach to the start codon of
the mRNA.

Releasing of the other initiation factors the larger ribosome subunit binds to the smaller
subunit which produces a fully functional ribosome. Following the formation of a fully
functional ribosome, the A site can again accept another aminoacyl-tRNA molecule.

By the end of the initiation phase, the initiation complex that is formed consists of both
ribosomal subunits (the large and smaller subunits), the mRNA, as well as the tRNA
that also carries N-formyl-methionine.

Elongation

The second phase of translation is known as elongation and is characterized by the

elongation of the polypeptide chain. Here, the ribosome has a catalytic function.
The entire process can be divided into three main steps of the elongation that include:
aminoacyl-tRNA binding, formation of the peptide bond, as well as translocation.
During the first step of this cycle (aminoacyl-tRNA binding), an aminoacyl-tRNA that
corresponds to the second codon binds to the A site (aminoacyl site) through the codon-
anticodon interaction.

Here, methionine that came with the initiation factor. tRna is released from the ribosome
when ribosome releasing factor binds onto the A site along with the initiator tRNA
during the initiation phase is the first amino acid. Binding of the aminoacyl-tRNA is
promoted by GTP and the elongation factor. The three join to form a complex
aminoacyl-tRNA which results in the hydrolysis of GTP. In turn, the elongation factor
is released.

The released molecule can then promote the binding of another tRNA to the ribosome
once it has regenerated. This occurs when elongation factor binds and replaces the GDP
on another elongation factor. The elongation factor are then replaced by GTP resulting
in the formation of a newly regenerated elongation factor.

The third step of the cycle (translocation) is characterized by binding of the elongation
complex and GTP to the ribosome. Here, hydrolysis of the GTP results in the production
of GDP and a phosphate while the release of the elongation factor frees it to bind GTP
in preparation of another elongation cycle.

With the deacylated tRNA moving from the P site to the E site and the dipeptidyl tRNA
from the A to the P site, the site remains empty and thus free to accept another
aminoacyltRNA. An amino acid is continually added to the C terminal end of the
polypeptide as it grows in length for each of the codons as the peptidyl-tRNA moves to
and from the P and A sites.

Termination

During elongation, the tRNA continually moves forward from the P to the A site as it
brings the next amino acid to be added onto the previous chain (chain that started with
a methionine). This process continues until a stop codon in the mRNA enters the A site
thus stopping the cycle from continuing. There are three types of stop codon which
include; UAA, UAG, and UGA.
The last phase of the translation process is known as termination and is the point at
which the process ends. Having entered the A site, the stop codon prevents tRNA from
binding.

One of the release factors binds to the codons causing enzyme (peptidyl transferase)
responsible for peptide bonds to release a water molecule into the last amino acid on
the chain which causes the peptide and tRNA attached to the P site to be hydrolyzed.
As a result, the newly formed chain is separated from tRNA and leaves the ribosome.
mRNA is released. tRNA is released from the ribosome when ribosome releasing factor
binds onto the A site. Release factors bind to the stop codon given that no tRNA have
anticodon for the stop codon in prokaryotes.

Translation in Eukaryotes
As is the case in prokaryotes, translation is the process through which a sequence of
mRNA is translated into polypeptides during protein synthesis.

As mentioned, transcription and translation processes occur in the cytoplasm in

prokaryotes (and can even occur at the same time). However, in eukaryotes, the
nucleus membrane separates the ribosome located in the cytoplasm from the
transcription process that takes place in the nucleus. For this reason, translation starts
when transcription ends and the mRNA is transported to the cytoplasm.

In eukaryotic organisms, translation also occurs in three phases that include

initiation, elongation, and termination.

Initiation

During the initiation phase, the smaller ribosomal subunit forms a complex with
three initiation factors. Here, however, the smaller ribosomal subunit is 40S
compared to the much smaller 30S in prokaryotes. Binding of these initiation factors
to the ribosomal subunit produce the preinitiation complex that in turn joins the
initiation factor and tRNA.

Ultimately, this complex binds the mRNA to form the initiation complex. As is the
case in prokaryotes, the small ribosomal subunit moves along the untranslated region
of the mRNA as it scans for the start codon (in most cases, the first AUG serves as
the start codon in eukaryotes).

Once the start codon is recognized, the larger ribosome (60S) subunit is recruited to
the complex which results in the formation of a fully functional ribosome (this is an
energy-dependent process that involves GTP hydrolysis and ultimately produces an
80S ribosome). Once a fully functional ribosome is formed, the initiation factors are
released. At the end of the initiation factor, the initiator tRNA met is located at the
P site while the A site remains vacant.
Elongation

This is the second phase of translation and involves the synthesis of polypeptide.
While the elongation process in eukaryotes is similar to that of eukaryotes. Here, the
elongation factor proteins has three main functions.

The first function of these proteins (elongation factor proteins) is to recruit the
charged tRNAs to the A site. In addition, they play an important role in forming a
peptide bond between the amino acids as well as translocation of the ribosome along
the mRNA.
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As the ribosome moves along the mRNA, elongation factors promote the peptide
bonds between amino acids located at on tRNA (at the A site) and the carboxyl group
of the amino group that is located on the tRNA of the P site.

Here, peptidyl transferase serves to catalyze the reaction. The amino acid associated
with the tRNA on the P site is then linked to the growing polypeptide chain which
allows the chain to continue growing in length. This process allows the ribosome to
continue moving along the mRNA as the polypeptide chain continues growing
before it stops at the termination phase.

Termination

This is the last phase of the translation process. It occurs when the ribosome arrives
at the nonsense codon of the mRNA where the tRNA has no complementary
anticodon. Once the nonsense codon is identified by release factors, the amino acid
at the P site is detached from the tRNA which frees the polypeptide.
On the other hand, the ribosome is not only dissociated from the mRNA, but also
into the two subunits (small and large ribosomal subunits) which allows them to
enter the initiation phase in another translation process.