Anda di halaman 1dari 39

MEKANISME EKSPRESI GEN

TERDARI ATAS DUA TAHAP


 Transkripsi

 Translasi
BATASAN TRANSKRIPS DAN TRANSLASI

 Transkripsi
◦ Adalah sintesis RNA di bawah arahan DNA
◦ Menghasilkan messenger RNA (mRNA) atau RNA
duta

 Translasi
◦ Adalah sintesis polipeptida yang terjadi di bawah
arahan mRNA
◦ Terjadi dalam ribosom
 Pada Prokariota
◦ Transkripsi dan translasi terjadi bersamaan (translasi
mRNA dimulai saat transkripsi masih berlangsung)

DNA
TRANSCRIPTION

mRNA
Ribosome

TRANSLATION

Polypeptide

(a) Prokaryotic cell. In a cell lacking a nucleus, mRNA


produced by transcription is immediately translated
Figure 17.3a without additional processing.
 Pada Eukariota
◦ Transkrip RNA atau transkrip primer atau pre-mRNA
dimodifikasi sebelum menjadi mRNA
Nuclear
envelope

TRANSCRIPTION DNA

Pre-mRNA
RNA PROCESSING

mRNA

Ribosome

TRANSLATION
(b) Eukaryotic cell. The nucleus provides a separate
Polypeptide compartment for transcription. The original RNA
transcript, called pre-mRNA, is processed in various
ways before leaving the nucleus as mRNA.
Figure 17.3b
 KOMPONEN TRANSKRIPSI

 DNA template
 Promoter pada DNA template
 Nukleotida
 RNA Polimerase

BIODAS II Dept. Biologi FST UNAIR


 PROMOTER

 URUTAN NUKLEOTIDA TERTENTU YANG


TERLETAK SEBELUM SEGMEN GEN
 FUNGSINYA UNTUK “TANDA PENGENAL”
GEN YANG AKAN DITRANSKRIP
 Dikenali oleh RNA polimerase
 Salah satu variannya (Pribnow Box) terdapat pada
posisi -10 dengan urutan yang dikenal dengan TATA
BOX sebab banyak mengandung T dan A
 Urutan lain posisinya pada -35

BIODAS II Dept. Biologi FST UNAIR


 ARN POLIMERASE
 Merupakan enzim yang tersusun dari 7000 molekul.
 Yang terlibat secara aktif pada saat btranskripsi hanya sekitar
2000 – 5000 molekul.
 Tersusun dari 5 subunit polipeptida: α, β, β’, ω dan σ
 Struktur aktifnya α2ββ’ωσ disebut holoenzim
 Struktur α2ββ’ω disebut core enzim
 Subunit σ (sigma) berungsi dalam pengenalam promoter.
 Subunit α berfungsi dalam menyusun enzim dan
menghubungkan dengan protein regulator.
 Subunit β berfungsi sebagai katalisator
 Subunit β’ berfungsi dalam “mengikat” DNA
 Subunit ω berperan dalam mengumpulkan enzim dan mengatur
ekspresi gen.

BIODAS II Dept. Biologi FST UNAIR


TAHAPAN TRANSKRIPSI

1. Inisiasi
2. Elongasi
3. Terminasi
INISIASI (1)
INISIASI
dimulai ketika subunit σ dari holoenzim ARN
polimerase
1 menempel pada promoter.

Daerah promoter terletak di sebelah hulu 5’ dari


gen yang ditranskrip.

Holoenzim akan menempel pada sekitar 60 pb


nukleotida heliks. 40 pb sebelum hulu (titik awal
transkripsi)

Daerah yang ditempeli holoenzim akan mengurai


(10 – 20pb).
INISIASI (2)
Daerah yang ditempeli holoenzim akan mengurai
(10 – 20pb) sehingga terbentuk ikatan promoter
terbuka.1

Setelah terbentuk ikatan promoter terbuka,


subunit σ berdesosiasi sehingga holoenzim
menjadi core enzim (subunit α, β, dan ω saja)

Pada saat bersamaan dua nukleotida dapat


berpasangan dengan pita template pada posisi +1
dan +2.

Ikatan fosfodiester pertama dari molekul ARN


terbentuk. Tahap inisiasi selesai.
ELONGASI

Kelanjutan dari tahap inisiasi


Polimerasi
1 terus bergerak maju, mengurai heliks,
sehingga haliks menjadi lurus (antara sekitar 17
pasang ARN-ADN).

Di daerah yang terutrai (gelembung DNA) 8-9


nukleotida ditambahkan pada ujung 3’, sedangkan
ujung 5’ dikeluarkan dari polimerase.
TERMINASI

TIDAK TERJADI SECARA RANDOM


Transkrip1
berakhir pada urutan tertentu yang
panjang sekitar 40 pb dan kaya akan G dan C.
Berakhir pada DNA palindrom (dapat
berpasangan dengan dirinya sendiri membentuk
struktur palindrom/”perempatan” yang diikuti 5 –
10 A atau U pada ARN.mekanisme penutupan ada
dua. Memerlukan prtin dan tidak memerlukan
protein.

Pada segmen penutup terdapat rho dependent


terminator.
ARN DILEPAS DARI DNA DAN POLIMERASE
BERDESOSIASI.
DUA TIPE TERMINASI

Mekanisme penutupan ada dua. Memerlukan prtin


dan tidak1 memerlukan protein.

1. Sada segmen terminator sepanjang sekitar 40


asang basa dan kaya akan GC. Segmen ini
membentu loop dan diikuti segmen yang
tersusun dari u sebanyak 8.
2. Memerlukan protein khusus rho dependent
terminator. Pada tipe ini ada segmen
sepanjang 40-60 nukleotida yang kaya akan U
dan miskin akan G. Protein ini akan mengikat
rantai RNA pada sisi celah yang terletak di
hulu RNA polimerase.
Sintesis Transkrip RNA
 Tahapan transkripsi
Promoter

◦ Inisiasi
Transcription unit
5 3
3 5
DNA

◦ Elongasi
Start point
RNA polymerase 1 Initiation. After RNA polymerase binds to
the promoter, the DNA strands unwind, and

◦ Terminasi
the polymerase initiates RNA synthesis at the
start point on the template strand.
5 3
3 5
Template strand of
Unwound RNA DNA
DNA transcript
2 Elongation. The polymerase moves downstream, unwinding the
DNA and elongating the RNA transcript 5  3 . In the wake of
Rewound transcription, the DNA strands re-form a double helix.

RNA
5 3
3 3 5
5

RNA
transcript
3 Termination. Eventually, the RNA
transcript is released, and the
polymerase detaches from the DNA.

5 3
3 5

5 3
Completed RNA
transcript
Pengikatan RNA Polimerase dan Inisiasi Transkripsi
TRANSCRIPTION DNA
1 Eukaryotic promoters

RNA PROCESSING Pre-mRNA

mRNA

 Promotor : sekuen DNA TRANSLATION Ribosome

Polypeptide

Promoter

tempat melekatnya RNA 5


3
T A T A A A A
A T A T T T T
3
5

polimerase dan tempat inisiasi TATA box Start point Template


DNA strand

untuk sintesis RNA 2 Several transcription


factors

Transcription
factors

Faktor Transkripsi
5 3

 3
3 Additional transcription
5

factors

◦ Membantu RNA polimerase


untuk mengenali sekuen
promoter pada Eukariota RNA polymerase II
Transcription factors

5 3
3 5 5

RNA transcript

Transcription initiation complex


Figure 17.8
Elongation Non-template
strand of DNA
RNA nucleotides

RNA
polymerase

T C C A A
A
3
3 end

A uE CG C A
5

T A G G T
T

Direction of transcription
5 Template
(“downstream”)
strand of DNA

Newly made
RNA
 Konsep 3: SEL EUKARIOTIK MEMODIFIKASI
RNA SETELAH TRANSKRIPSI

 Enzim di inti sel pada eukariotik


◦ Modifikasi pre-mRNA dengan cara tertentu
sebelum pesan genetik dikirim ke sitoplasma

BIODAS II Dept. Biologi FST UNAIR


Pengubahan ujung mRNA
 Setiap ujung molekul pre-mRNA dimodifikasi
dengan cara tertentu
◦ Ujung 5 dimodifikasi dengan penambahan
nukeotida tudung (cap)
◦ Ujung 3 dengan penambahan ekor poli-A

A modified guanine nucleotide 50 to 250 adenine nucleotides


added to the 5 end added to the 3 end
TRANSCRIPTION DNA

RNA PROCESSING Pre-mRNA Protein-coding segment Polyadenylation signal


5
3
mRNA
G P P P AAUAAA AAA…AAA
Ribosome
TRANSLATION
Start codon Stop codon
5 Cap 5 UTR 3 UTR Poly-A tail
Polypeptide

Figure 17.9
BIODAS II Dept. Biologi FST UNAIR
Split Genes dan RNA Splicing
 Penyambungan RNA / RNA splicing
◦ Memindahkan intron dan menggabungkan exon

5 Exon Intron Exon Intron Exon 3


TRANSCRIPTION DNA Pre-mRNA 5 Cap Poly-A tail
1 30 31 104 105 146
RNA PROCESSING Pre-mRNA

mRNA Coding Introns cut out and


segment exons spliced together
Ribosome
TRANSLATION

Polypeptide
mRNA 5 Cap Poly-A tail
1 146
3 UTR 3 UTR

Figure 17.10
BIODAS II Dept. Biologi FST UNAIR
 Dilakukan oleh spliceosome, yaitu enzim yang berfungsi
untuk memindahkan intron dan menggabungkan exon

RNA transcript (pre-mRNA)


5
Exon 1 Intron Exon 2

Protein
1 Other proteins
snRNA

snRNPs
Spliceosome

2 5

Spliceosome
components
Cut-out
intron
3
mRNA
Figure 17.11 5
Exon 1 Exon 2
BIODAS II Dept. Biologi FST UNAIR
 Protein sering memiliki arsitektur modular
◦ Terdiri dari daerah struktural dan fungsional yang
disebut domain
◦ Kode exon akan berbeda untuk domain yang
berbeda pada protein
Gene
DNA
Exon 1 Intron Exon 2 Intron Exon 3

Transcription
RNA processing

Translation

Domain 3

Domain 2

Domain 1

Figure 17.12
Polypeptide
BIODAS II Dept. Biologi FST UNAIR
 Konsep 4: TRANSLASI ADALAH SINTESIS
POLIPEPTIDA YANG DIARAHKAN OLEH RNA

Komponen Molekular Translasi


 Sel mentranslasi pesan mRNA menjadi
protein dengan bantuan RNA transfer (tRNA)

BIODAS II Dept. Biologi FST UNAIR


 Konsep Dasar Translasi
TRANSCRIPTION DNA

mRNA
Ribosome
TRANSLATION
Polypeptide

Amino
Polypeptide acids

tRNA with
amino acid
attached
Ribosome

Gly

tRNA

Anticodon
A A A
U G G U U U G G C

5 Codons 3
Figure 17.13 mRNA
BIODAS II Dept. Biologi FST UNAIR
Struktur dan Funsi RNA Transfer
 Molekul tRNA
◦ Tersusun dari untai RNA tunggal yang panjangnya
A
C
hanya
sekitar 80 nukleotida C
3
◦ Berbentuk L Amino acid
attachment site
A
C
C
A 5
C G
G C
C G
U G
U A
A U
U C A U
* C A C AG U A G *
A
G * C U C *
C G U G U * C G A G G
* * U C *
A G G
* G AG C
(a) Two-dimensional structure. The four base-paired regions and three G C Hydrogen
loops are characteristic of all tRNAs, as is the base sequence of the U A bonds
* G
amino acid attachment site at the 3 end. The anticodon triplet is unique A
to each tRNA type. (The asterisks mark bases that have been chemically A * C
modified, a characteristic of tRNA.) * U
A G
A

Figure 17.14a Anticodon


BIODAS II Dept. Biologi FST UNAIR
5 Amino acid
attachment site
3

Hydrogen
bonds

A A G

3 5
Anticodon
Anticodon
(c) Symbol used
in this book
(b) Three-dimensional structure

Figure 17.14b
BIODAS II Dept. Biologi FST UNAIR
 Enzim aminoacyl-tRNA synthetase
◦ Menggabungkan masing-masing asam amino ke tRNA
yang benar

Amino acid Aminoacyl-tRNA


synthetase (enzyme)

P P P Adenosine
1 Active site binds the
amino acid and ATP.
ATP

2 ATP loses two P groups


and joins amino acid as AMP.
P Adenosine

Pyrophosphate P Pi

Pi
Pi
Phosphates
tRNA
3 Appropriate
tRNA covalently
Bonds to amino
Acid, displacing P Adenosine
AMP. AMP
4 Activated amino acid
is released by the enzyme.

Aminoacyl tRNA
(an “activated
Figure 17.15 amino acid”)

BIODAS II Dept. Biologi FST UNAIR


Ribosom
◦ Menfasilitasi ikatan antikodon tRNA dengan kodon mRNA
selama sintesis protein
TRANSCRIPTION DNA

mRNA
Ribosome
TRANSLATION

Subunit ribosomal
Polypeptide

 Growing
Exit tunnel

polypeptide

◦ Disusun oleh protein tRNA


molecules
dan molekul RNA yang Large
subunit

dinamakan RNA E
P
A

ribosomal atau rRNA Small


subunit

5
mRNA 3

(a)
Computer model of functioning ribosome. This is a model of a bacterial
ribosome, showing its overall shape. The eukaryotic ribosome is roughly similar. A
ribosomal subunit is an aggregate of ribosomal RNA molecules and proteins.

Figure 17.16a
BIODAS II Dept. Biologi FST UNAIR
 Ribosom mempunyai 3 sisi pengikatan
(binding sites) tRNA
◦ Sisi P P site (Peptidyl-tRNA
binding site)
◦ Sisi A A site (Aminoacyl-
tRNA binding site)

◦ Sisi E E site
(Exit site)

Large
subunit

E P A

mRNA
binding site
Small
subunit

(b) Schematic model showing binding sites. A ribosome has an mRNA binding site
and three tRNA binding sites, known as the A, P, and E sites. This schematic
Figure 17.16b ribosome will appear in later diagrams.

BIODAS II Dept. Biologi FST UNAIR


Amino end Growing polypeptide

Next amino acid


to be added to
polypeptide chain

tRNA

mRNA 3

Codons
5

(c) Schematic model with mRNA and tRNA. A tRNA fits into a binding site when its anticodon base-pairs
with an mRNA codon. The P site holds the tRNA attached to the growing polypeptide. The A site holds
the tRNA carrying the next amino acid to be added to the polypeptide chain. Discharged tRNA leaves via
the E site.

Figure 17.16c

BIODAS II Dept. Biologi FST UNAIR


Pembentukan Polipeptida
 Kita dapat membagi translasi menjadi 3
tahapan
◦ Inisiasi
◦ Elongasi
◦ Terminasi

BIODAS II Dept. Biologi FST UNAIR


Asosiasi Ribosom dan Inisiasi Translasi
 Tahap inisiasi translasi
◦ Menyatukan mRNA, tRNA yang membawa asam amino
pertama dari polipeptida, dan dua subunit ribosom
Large
ribosomal
P site subunit
3 U A C 5
5 A U G 3

Initiator tRNA
GTP GDP
E A
mRNA
5 3 5 3
Start codon

mRNA binding site Small Translation initiation complex


ribosomal
subunit

1 2
A small ribosomal subunit binds to a molecule of The arrival of a large ribosomal subunit completes
mRNA. In a prokaryotic cell, the mRNA binding site the initiation complex. Proteins called initiation
on this subunit recognizes a specific nucleotide factors (not shown) are required to bring all the
sequence on the mRNA just upstream of the start translation components together. GTP provides
codon. An initiator tRNA, with the anticodon UAC, the energy for the assembly. The initiator tRNA is
base-pairs with the start codon, AUG. This tRNA in the P site; the A site is available to the tRNA
carries the amino acid methionine (Met). bearing the next amino acid.
Figure 17.17 BIODAS II Dept. Biologi FST UNAIR
Elongasi Translasi
◦ Asam amino ditambahkan satu per satu ke asam amino
sebelumnya

1 Codon recognition. The anticodon


TRANSCRIPTION DNA
Amino end of an incoming aminoacyl tRNA
mRNA
of polypeptide base-pairs with the complementary
Ribosome
TRANSLATION mRNA codon in the A site. Hydrolysis
Polypeptide
of GTP increases the accuracy and
E efficiency of this step.
mRNA 3
Ribosome ready for P A
5 site site
next aminoacyl tRNA
2 GTP
2 GDP

E E

P A P A

2 Peptide bond formation. An


GDP rRNA molecule of the large
3 Translocation. The ribosome GTP subunit catalyzes the formation
translocates the tRNA in the A
of a peptide bond between the
site to the P site. The empty tRNA
new amino acid in the A site and
in the P site is moved to the E site, E
the carboxyl end of the growing
where it is released. The mRNA
polypeptide in the P site. This step
moves along with its bound tRNAs, P A attaches the polypeptide to the
bringing the next codon to be
tRNA in the A site.
Figure 17.18 translated into the A site.
BIODAS II Dept. Biologi FST UNAIR
Terminasi Translasi
 Tahap akhir translasi adalah ketika ribosom
sampai kodon stop pada mRNA

Release
factor
Free
polypeptide

5
3 3
3
5 5
Stop codon
(UAG, UAA, or UGA)
1 When a ribosome reaches a stop 2 The release factor hydrolyzes 3 The two ribosomal subunits
codon on mRNA, the A site of the the bond between the tRNA in and the other components of
ribosome accepts a protein called the P site and the last amino the assembly dissociate.
a release factor instead of tRNA. acid of the polypeptide chain.
The polypeptide is thus freed
from the ribosome.
Figure 17.19
BIODAS II Dept. Biologi FST UNAIR
Poliribosom
 Sejumlah ribosom dapat mentranslasi satu molekul
mRNA tunggal secara simultan. Hanya pada sel
prokariota Growing
Completed
polypeptide
polypeptides

Incoming
ribosomal
subunits
Start of End of
mRNA mRNA
(5 end) (3 end)
(a) An mRNA molecule is generally translated simultaneously
by several ribosomes in clusters called polyribosomes.

Ribosomes
mRNA

0.1 µm
(b) This micrograph shows a large polyribosome in a prokaryotic
Figure 17.20a, b cell (TEM).
BIODAS II Dept. Biologi FST UNAIR
Protein Fungsional

 Rantai polipeptida /protein menjalani


modifikasi setelah proses translasi

 Protein dimodifikasi
- untuk membentuk molekul tiga dimensi / folding
(pelipatan)
- Untuk dapat ditranfer ke lokasi yang membutuhkan

BIODAS II Dept. Biologi FST UNAIR


 Protein dibutuhkan dalam sistem
endomembran (contoh: enzim lisosim yang ada
di organel lisosom) atau disekresikan (contoh:
enzim pencernaan seperti amilase).

 Protein harus ditransport ke retikulum


endoplasma (ER)
Signal peptida pada ribosom binding pada
signal-recognition particle (SRP). Kompleks
keduanya binding pada protein reseptor SRP di
ER

BIODAS II Dept. Biologi FST UNAIR


 Mekanisme signal untuk protein target pada ER

1 Polypeptide 2 An SRP binds 3 The SRP binds to a 4 The SRP leaves, and 5 The signal- 6 The rest of
synthesis begins to the signal receptor protein in the ER the polypeptide resumes cleaving the completed
on a free peptide, halting membrane. This receptor growing, meanwhile enzyme polypeptide leaves
ribosome in synthesis is part of a protein complex translocating across the cuts off the the ribosome and
the cytosol. momentarily. (a translocation complex) membrane. (The signal signal peptide. folds into its final
that has a membrane pore peptide stays attached conformation.
and a signal-cleaving enzyme. to the membrane.)

Ribosome

mRNA
Signal
peptide ER
membrane
Signal
Signal-
peptide
recognition Protein
removed
particle
(SRP) SRP
receptor
CYTOSOL protein

Translocation
ERLUMEN
complex

Figure 17.21
 Ringkasan transkripsi dan translasi pada sel eukariotik
TRANSCRIPTION DNA
1 RNA is transcribed
from a DNA template.
3

5 RNA RNA
transcript polymerase
RNA PROCESSING Exon
2 In eukaryotes, the
RNA transcript
RNA transcript (pre- (pre-mRNA)
mRNA) is spliced and
Intron
modified to produce
mRNA, which moves Aminoacyl-tRNA
from the nucleus to the synthetase
cytoplasm. NUCLEUS

Amino
FORMATION OF acid
INITIATION COMPLEX AMINO ACID ACTIVATION
CYTOPLASM tRNA
3 After leaving the 4 Each amino acid
nucleus, mRNA attaches attaches to its proper tRNA
to the ribosome. with the help of a specific
enzyme and ATP.
mRNA Growing
polypeptide
Activated
amino acid

Ribosomal
subunits

5
TRANSLATION
5 A succession of tRNAs
E A add their amino acids to
AAA Anticodon the polypeptide chain
as the mRNA is moved
UGGUU UA U G
through the ribosome
one codon at a time.
Codon (When completed, the
polypeptide is released
Ribosome
Figure 17.26 from the ribosome.)

BIODAS II Dept. Biologi FST UNAIR


39