Kuliah 9.

Translation
Dwi Suryanto
Program Studi Biologi
Universitas Sumatera Utara
dwisuryanto@usu.ac.id
Untuk kalangan sendiri dan diambil dari banyak sumber WA: 082165131177
 Post-Transcription in Eukaryote
After transcription, eukaryotic pre-mRNAs must undergo
several processing steps before they can be translated.
Eukaryotic (and prokaryotic) tRNAs and rRNAs also undergo
processing before they can function as components in the
protein synthesis machinery.
Post transcription editing
mRNA Processing
The eukaryotic pre-mRNA undergoes extensive processing before it is
ready to be translated. The additional steps involved in eukaryotic mRNA
maturation create a molecule with a much longer half-life than a
prokaryotic mRNA. Eukaryotic mRNAs last for several hours, whereas the
typical E. coli mRNA lasts no more than five seconds.
Pre-mRNAs are first coated in RNA-stabilizing proteins; these protect the
pre-mRNA from degradation while it is processed and exported out of the
nucleus. The three most important steps of pre-mRNA processing are the
addition of stabilizing and signaling factors at the 5′ and 3′ ends of the
molecule, and the removal of intervening sequences that do not specify
the appropriate amino acids. In rare cases, the mRNA transcript can be
“edited” after it is transcribed.
5′ Capping
While the pre-mRNA is still being synthesized, a 7-methylguanosine
cap is added to the 5′ end of the growing transcript by a phosphate
linkage. This moiety (functional group) protects the nascent mRNA from
degradation. In addition, factors involved in protein synthesis recognize
the cap to help initiate translation by ribosomes.
3′ Poly-A Tail
Once elongation is complete, the pre-mRNA is cleaved by an endonuclease
between an AAUAAA consensus sequence and a GU-rich sequence,
leaving the AAUAAA sequence on the pre-mRNA. An enzyme called poly-A
polymerase then adds a string of approximately 200 A residues, called
the poly-A tail. This modification further protects the pre-mRNA from
degradation and signals the export of the cellular factors that the
transcript needs to the cytoplasm.
Pre-mRNA Splicing
Eukaryotic genes are composed of exons, which correspond
to protein-coding sequences (ex-on signifies that they
are expressed), and intervening sequences
called introns (intron denotes their intervening role), which
may be involved in gene regulation but are removed from the
pre-mRNA during processing. Intron sequences in mRNA do
not encode functional proteins.
The discovery of introns came as a surprise to researchers in the 1970s
who expected that pre-mRNAs would specify protein sequences without
further processing, as they had observed in prokaryotes. The genes of
higher eukaryotes very often contain one or more introns. These regions
may correspond to regulatory sequences; however, the biological
significance of having many introns or having very long introns in a gene
is unclear. It is possible that introns slow down gene expression because
it takes longer to transcribe pre-mRNAs with lots of introns. Alternatively,
introns may be nonfunctional sequence remnants left over from the fusion
of ancient genes throughout evolution. This is supported by the fact that
separate exons often encode separate protein subunits or domains. For
the most part, the sequences of introns can be mutated without ultimately
affecting the protein product.
All of a pre-mRNA’s introns must be completely and precisely
removed before protein synthesis. If the process errs by even
a single nucleotide, the reading frame of the rejoined exons
would shift, and the resulting protein would be dysfunctional.
The process of removing introns and reconnecting exons is
called splicing. Introns are removed and degraded while the
pre-mRNA is still in the nucleus. Splicing occurs by a
sequence-specific mechanism that ensures introns will be
removed and exons rejoined with the accuracy and precision
of a single nucleotide. The splicing of pre-mRNAs is
conducted by complexes of proteins and RNA molecules
called spliceosomes.
 Universal Genetic Code
DNA is considered a universal genetic code because every known
living organism has genes made of DNA. Bacteria, fungi, cats, plants,
and you: every organism uses DNA to store genetic information. All
organisms also use DNA to transcribe RNA, and then they translate
that RNA into proteins.
The genetic code
is the set of rules
used by living cells
to translate
information
encoded within
genetic material
into proteins.
Universal genetic code
Meskipun universal, ada beberapa kodon tidak sama dalam hal
terjemahan asam aminonya.
 Prokaryote Translation
• Translasi: sintesis suatu polipeptida menggunakan
infomasi genetik (kode genetik) yang disandi dalam satu
molekul mRNA.
• Terjadi perubahan “bahasa” dari nukleotida menjadi asam
amino.
• Terdapat 3 jenis RNA yang terlibat: mRNA, rRNA, dan tRNA
Messenger ribonucleic acid is a single-stranded molecule of RNA
that corresponds to the genetic sequence of a gene, and is read by a
ribosome in the process of synthesizing a protein.
Ribosomal ribonucleic acid is a
type of non-coding RNA which is
the primary component of
ribosomes, essential to all cells.
rRNA is a ribozyme which carries
out protein synthesis in
ribosomes. Within the ribosome,
the rRNA molecules direct the
catalytic steps of protein
synthesis — the stitching together
of amino acids to make a protein
molecule. In fact, rRNA is
sometimes called a ribozyme or
catalytic RNA to reflect this
function.
A ribosome is an
intercellular structure
made of both RNA and
protein, and it is the
site of protein synthesis
in the cell. The
ribosome reads the
messenger RNA (mRNA)
sequence and translates
that genetic code into a
specified string of amino
acids, which grow into
long chains that fold to
form proteins.
Perbandingan ribosom Prokaryota dengan ribosom Eukaryota
Transfer RNA is an adaptor molecule composed of RNA, typically
76 to 90 nucleotides in length, that serves as the physical link
between the mRNA and the amino acid sequence of proteins.
Transfer RNA does this by carrying an amino acid to the protein
synthesizing machinery of a cell called the ribosome.

Suatu jenis molekul RNA yang berfungsi sebagai penerjemah

antara “bahasa” asam nukleat dan protein dengan cara
membawa asam amino tertentu dan mengenali kodon yang
sesuai dalam mRNA.
Molekul tRNA
a. Inisiation of translation
Initiation of translation in bacteria involves the assembly of
the components of the translation system, which are: the two
ribosomal subunits (50S and 30S subunits); the mature
mRNA to be translated; the tRNA charged with N-
formylmethionine (the first amino acid in the nascent
peptide); guanosine triphosphate (GTP) as a source of energy,
and the three prokaryotic initiation factors IF1, IF2, and IF3,
which help the assembly of the initiation complex. Variations
in the mechanism can be anticipated.
The ribosome has three active sites: the A site, the P site, and
the E site. The A site is the point of entry for the aminoacyl
tRNA (except for the first aminoacyl tRNA, which enters at the
P site). The P site is where the peptidyl tRNA is formed in the
ribosome. And the E site which is the exit site of the now
uncharged tRNA after it gives its amino acid to the growing
peptide chain.
The selection of an initiation site (usually an AUG codon) depends on the
interaction between the 30S subunit and the mRNA template. The 30S
subunit binds to the mRNA template at a purine-rich region (the Shine-
Dalgarno sequence) upstream of the AUG initiation codon. The Shine-
Dalgarno sequence is complementary to a pyrimidine rich region on the 16S
rRNA component of the 30S subunit. This sequence has been evolutionarily
conserved and plays a major role in the microbial world we know today.
During the formation of the initiation complex, these
complementary nucleotide sequences pair to form a double
stranded RNA structure that binds the mRNA to the ribosome
in such a way that the initiation codon is placed at the P site.
b. Elongation of translation
The process of polypeptide chain elongation is basically the
same in both prokaryotes and eukaryotes. The addition of
each amino acid to the growing polypeptide occurs in three
steps:
• Step 1: binding of an aminoacyl-tRNA to the A site of the
ribosome
• Step 2: transfer of the growing polypeptide chain from the
tRNA in the P site to the tRNA in the A site by the formation
of a new peptide bond,
• Step 3: translocation of the ribosome along the mRNA to
position the next codon in the A site.
Elongation of translation
b. Termination of translation
• Terminasi terjadi ketika kodon stop masuk ke dalam
ribosom, sementara tRNA dengan antikodon yang sesuai
tidak membawa asam amino.
Translation
Faktor (protein) yang terlibat dalam translasi prokaryote
 Eukaryote Translation
• The overall mechanism of protein synthesis in eukaryotes is
basically the same as in prokaryotes.
• However, there are some significant differences:
• Whereas a prokaryotic ribosome has a sedimentation coefficient
of the 70S and subunits of 30S and 50S, a eukaryotic ribosome
has a sedimentation coefficient of 80S with subunits of 40S and
60S.
• The composition of eukaryotic ribosomal subunits is also more
complex than prokaryotic subunits but the function of each
subunit is essentially the same as in prokaryotes.
• The composition of eukaryotic ribosomal subunits is also more
complex than prokaryotic subunits but the function of each
subunit is essentially the same as in prokaryotes.
• In eukaryotes, each mRNA is monocistronic that is, discounting
any subsequent post-translational cleavage reactions that may
occur; the mRNA encodes a single protein. In prokaryotes, many
mRNAs are polycistronic that is they encode several proteins.
Each coding sequence in a prokaryotic mRNA has its own
initiation and termination codons.
• Initiation of protein synthesis in eukaryotes requires at least
nine distinct eukaryotic initiation factors (eIFs) compared with
the three initiation factors (IFs) in prokaryotes.
• In eukaryotes, the initiating amino acid is methionine, not N-
formylmethionine as in prokaryotes.
• As in prokaryotes, a special initiator tRNA is required for
initiation and is distinct from the tRNA that recognizes and binds
to codons for methionine at internal positions in the mRNA.
When charged with methionine ready to begin initiation, this is
known as Met-tRNAimet
• The main difference between initiation of translation in
prokaryotes and eukaryotes is that in bacteria, a Shine–Dalgarno
sequence lies 5’ to the AUG initiation codon and is the binding
site for the 30S ribosomal subunit.
• In contrast, most eukaryotic mRNAs do not contain Shine–
Dalgarno sequences. Instead, a 40S ribosomal subunit attaches
at the 5’ end of the mRNA and moves downstream (i.e. in a 5’ to
3’ direction) until it finds the AUG initiation codon. This process
is called scanning.
• Prokaryotic translation requires no helicase, presumably
because protein synthesis in bacteria can start even as the
mRNA is still being synthesized whereas, in eukaryotes,
transcription in the nucleus and translation in the cytoplasm are
separate events that allow time for mRNA secondary structure to
form.
 Diskusi
“ Jika engkau punya satu apel dan aku punya satu apel lalu kita
pertukarkan maka kita masing-masing hanya punya satu apel, jika
engkau punya satu ide dan aku punya satu ide lalu kita pertukarkan
maka kita masing-masing punya dua ide” (George B. Shaw)
Terima Kasih