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Translation: From Messenger RNA To Protein

The document summarizes how genetic information encoded in DNA is transcribed into messenger RNA and then translated into proteins with the help of the ribosome. It describes the processes of transcription, where DNA is copied into mRNA, and translation, where the mRNA code is decoded by ribosomes along with tRNAs to produce a protein. It provides details on the structures and mechanisms involved in translation, including initiation, elongation, termination and factors that regulate translation.

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37 views

Translation: From Messenger RNA To Protein

The document summarizes how genetic information encoded in DNA is transcribed into messenger RNA and then translated into proteins with the help of the ribosome. It describes the processes of transcription, where DNA is copied into mRNA, and translation, where the mRNA code is decoded by ribosomes along with tRNAs to produce a protein. It provides details on the structures and mechanisms involved in translation, including initiation, elongation, termination and factors that regulate translation.

Uploaded by

bombertest1
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
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Translation: From messenger RNA to protein:

The information encoded in the DNA is transferred to


messenger RNA and then decoded by the ribosome to
produce proteins.
5’-ATGCCTAGGTACCTATGA-3’ DNA
3’-TACGGATCCATGGATACT-5’
Transcription
5’-AUGCCUAGGUACCUAUGA-3’ mRNA

decoded as

5’-AUG CCU AGG UAC CUA UGA-3’


Translation

N-MET-PRO-ARG-TYR-LEU-C Protein
Alanine tRNA
Generalized tRNA
Modified Bases
Found in tRNAs

= UH2
tRNAs are activated by amino-acyl tRNA synthetases
Structure of an amino acyl-tRNA synthetase bound to a tRNA
One mechanism for maintaining high fidelity of protein
synthesis is the high fidelity of aa-tRNA synthetases
Amino-acyl tRNA synthetases:
One synthetase for each amino acid
a single synthetase may recognize multiple tRNAs
for the same amino acid

Two classes of synthetase.


Different 3-dimensional structures
Differ in which side of the tRNA they recognize
and how they bind ATP
Class I - monomeric, acylates the 2’OH on the terminal ribose
Arg, Cys , Gln, Glu, Ile, Leu, Met, Trp Tyr, Val

Class II - dimeric, acylate the 3’OH on the terminal ribose


Ala, Asn, Asp, Gly, His, Lys, Phe, Ser, Pro, Thr
Two levels of control to ensure that the proper amino acid
is incorporated into protein: 1) Charging of the proper tRNA
2) Matching the
cognate tRNA to the
messenger RNA
Incorporation of amino acids into polypeptide chains I
Incorporation of amino acids into polypeptide chains II
Protein synthesis occurs on ribosomes
Protein synthesis occurs on ribosomes
and mitochondria
Ribosome Assembly

The proteins of each


ribosomal subunit
are organized around
rRNA molecules

16S rRNA
Ribosome Assembly: takes place largely in a specialized domain of
the nucleus, the nucleolus
In the nucleolus, RNA polymerase I transcribes the rDNA repeats
to produce a 45S RNA precursor

The 45S precursor


is processed
and cleaved into
mature rRNAs and
ribosomal proteins
then bind to generate
the large and small
ribosomal subunits
23S rRNA secondary structure
3D organization of the eukaryotic large subunit rRNA
Ribosomal Proteins decorate the surface of the ribosome

Large subunit. Grey = rRNA Gold = ribosomal proteins


Ribosomal proteins often have extensions that snake into
the core of the rRNA structure

Crystal structure of L19 L15 (yellow) positioned in a fragment


of the rRNA (red)
The ribosomal proteins are important for maintaining the
stability and integrity of the ribosome, but NOT for catalysis

ie. the ribosomal RNA acts as a ribozyme


The large and small subunits come together to form the ribosome

Mitochondrial
or Prokaryotic
Eukaryotic 60S subunit 80S ribosome 40S subunit
The association of the large and small subunits creates the
structural features on the ribosome that are essential for
protein synthesis

Three tRNA binding


sites:
A site = amino-acyl
tRNA binding site

P site = peptidyl-tRNA
binding site

E site = exit site


In addition to the APE sites there is an mRNA binding groove
that holds onto the message being translated
There is a tunnel through the large subunit that allows the
growing polypeptide chain to pass out of the ribosome
Peptide bond formation is catalyzed by the large subunit rRNA
Peptide bond formation is catalyzed by the large subunit rRNA
Incorporation of the correct amino acyl-tRNA is determined
by base-pairing interactions between the anticodon of the
tRNA and the messenger RNA
=EF-1 Proper reading of the
anticodon is the second
important quality control
step ensuring accurate
protein synthesis

Elongation factors
Introduce a two-step
“Kinetic proofreading”
A second elongation factor
EF-G or EF-2, drives the
translocation of the ribosome
along the mRNA

Together GTP hydrolysis


by EF-1 and EF-2 help drive
protein synthesis forward
Termination of translation
is triggered by stop codons

Release factor enters


the A site and triggers
hydrolysis the peptidyl-tRNA
bond leading to release of
the protein.
Release of the protein causes
the disassociation of the
ribosome into its constituent
subunits.
Release Factor is a molecular mimic of a tRNA

eRF1 tRNA
Initiation of Translation

Initiation is controlled differently in prokaryotic and


eukaryotic ribosomes

In prokaryotes a single transcript can give rise to multiple proteins


In prokaryotes, specific
sequences in the mRNA around
the AUG codon, called
Shine-Delgarno sequences,
are recognized by an intiation
complex consisting of a Met
amino-acyl tRNA, Initiation
Factors (IFs) and the small
ribosomal subunit
GTP hydrolysis by
IF2 coincident with
release of the IFs and
binding of the large
ribosomal subunit leads
to formation of a complete
ribosome,on the mRNA
and ready to translate.
Eukaryotic mRNAs have a distinct structure at the 5’ end
Structure of the 7-methyl guanosine cap

The 7me-G cap is required


for an mRNA to be
translated
In contrast, Eukaryotes
use a scanning mechanism
to intiate translation.

Recognition of the AUG


triggers GTP hydrolysis
by eIF-2
GTP hydrolysis by
eIF2 is a signal for
binding of the large
subunit and beginning
of translation
Messenger RNAs are translated on polyribosomes
Protein synthesis is often regulated at the
level of translation initiation
An example of control of specific mRNAs: regulation by iron (Fe):

Ferritin is a cytosolic iron binding protein expressed when


iron is abundant in the cell.

Transferrin receptor is a plasma membrane receptor important


for the import of iron into the cytosol.

They are coordinately regulated, in opposite directions, by


control of protein synthesis.
Regulation by iron (Fe):
There is also general control of translational initiation.

ie. all transcripts of the cell are effected (though the relative
effect differs between specific mRNAs)

Global downregulation or upregulation can occur in response


to various stimuli the most common are
1) Nutrient availability
low nutrient (amino acids/carbohydrate)
downregulates translation

2) Growth factor signals.


stimulation of cell division upregulates translation
General control of translational initiation is exerted through
two primary mechanisms.

Control of the phosphorylation of eIF2

Control of the phosphorylation of eIF4 binding proteins


Control of translation by eIF2 phosphorylation

Stimulated by
Amino acid deprivation
Control of translation by eIF4E availability

The 7MEG cap binding subunit of eIF4, eIF4E, is sequestered


by eIF4E binding protiens (4E-BPs). The binding of these
proteins is regulated by their phosphorylation state

Growth Nutrient
Factors Limitation
Nutritional
2
controls

Nutritional signals can control both the recognition of the mRNA


and loading of the 40S subunit.
Modification of the translation machinery is a common
feature of viral life cycles

e.g. Picornaviruses
Polio virus
Encephalomyocarditis virus

Picornaviruses have single stranded RNA genomes.


Poliovirus Life Cycle
The poliovirus genome is translated into a single,
large polyprotein that then auto-proteolyzes itself into
smaller proteins.

One of these proteins, viral protease 2A cleaves


the translation initiation factor eIF4G so that it
can no longer function as a bridge between the
methyl cap binding subunit and the 40S subunit
The consequence of this cleavage is that translation of
cellular mRNAs stops

But…the viral RNA is still translated due to the presence of


an internal ribosomal entry site (IRES). This acts like a
bacterial initiation site to allow Cap-independent initiation
from internal AUG codons.

What is X?
“X” is not a protein, as suggested
by the textbook model at right,
rather it is a structure in the mRNA
itself that can bind to the remaining
fragment of eIF4G
Some cellular mRNAs are also translated using IRESs

During G2/M phase of the cell cycle, translation is generally


downregulated by activation of 4E-BPs. Many proteins expressed
during this period bypass this control by using IRES elements
Ribosomal Frameshifting

Because translation
uses a triplet code,
there are three potential
reading frames in
each mRNA
As the ribosome translocates, it moves in three nucleotide
steps, ensuring that the frame defined by the AUG is used
throughout translation

If the ribosome moves 1 or 2 (or 4 or 5) nucleotides


this produces a frameshift
Many retroviruses induce ribosomal frameshifting in
the synthesis of viral proteins

e.g. HIV
Translation Inhibitors are important antibiotics

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