Anda di halaman 1dari 88

REGULASI DAN

EKSPRESI GEN
SUSANTI
EVY SETYOWATI
RAHARJA KUNCARA

RNA
Ada 3 tipe RNA:
RNAd membawa kode genetik ke ribosom
RNAr komponen utama ribosom
RNAt membawa asam amino ke ribosom

Rantai RNAd.

Struktur kimia urasil dan


ribosa pada RNA.

Struktur RNAt.

RNA ( Ribonukleic Acid)


Merupakan benang tunggal, tersusun dari gula
ribosa, Posfat dan Basa. Basa purin RNA terdiri
dari Adenin (A) dan Guanin (G) sedangkan basa
pirimidin terdiri dari Sitosin (S) dan Urasil (U).
Ditemukan di inti sel, mitokondria, kloroplast dan
didalam sitoplasma.
Berdasarkan fungsinya, RNA di bagi 3 macam :
1 RNA Duta (RNA-d) RNA Mesenjer
2
RNA Ribosom (RNA-r)
3
RNA transfer (RNA-t)

RNA d atau RNA-m.


Merupakan polinukleotida berbentuk
linier, disintesis didalam nukleus
melalui transkripsi oleh DNAyg
berfungsi sebagai penentu primer
dari protein yg akan dibentuk
(membawa kodon)
Merupakan rantai tunggal yang relatif
panjang yang berfungsi menentukan
spesifikasi urutan asam amino pada
rantai polipeptida.

RNA- r atau RNA ribosom


Merupakan penyusun ribosom
dengan bentuk yang belum
diketahui
Berfungsi sebagai adaptor
atau penyelaras dalam
sintesis protein
Sekitar 80%RNA didalam sel
adalah RNA-r.

RNA-t (RNA transfer)


Berperan dalam memanggil asam
amino spesifik,mengikatnya dan
kemudian mengangkutnya menuju
ribosom.
Pada satu bagiannya akan terikat
asam amino, sedang pada ujung
yang lain memiliki urutan basa
(anti kodon) yang nantinya akan
mencari pasangannya dari RNA-d
(kodon)

tRNA
The genetic code is read during translation
via adapter molecules, tRNAs, that have 3base anticodons complementary to
codons in mRNA.
"Wobble" during reading of the mRNA
allows some tRNAs to read multiple codons
that differ only in the 3rd base.
There are 61 codons specifying 20 amino
acids. Minimally 31 tRNAs are required for
translation, not counting the tRNA that
codes for chain initiation. Mammalian cells
produce more than 150 tRNAs.

RNA structure:
Most RNA molecules have
secondary structure,
consisting of stem & loop
domains.

A
:
U

U
:
A

A
:
U

stem

C
:
G

C UG
C
:
U
G
U
C U

loop

Double helical stem domains arise from


base pairing between complementary
stretches of bases within the same strand.
These stem structures are stabilized by
stacking interactions as well as base pairing,
as in DNA.
Loop domains occur where lack of
complementarity or the presence of

anticodon loop

tRNA

acceptor
stem

The cloverleaf model of tRNA


emphasizes the two major types of
secondary structure, stems & loops.
tRNAs typically include many modified
bases, particularly in loop domains.

RNA tertiary structure depends on


interactions of bases at distant sites.
These interactions generally involve nonstandard base pairing and/or interactions
involving three or more bases.
Unpaired adenosines (not involved in
conventional base pairing) predominate
in participating in non-standard
interactions that stabilize tertiary RNA
structures.
tRNAs have an L-shaped tertiary structure.

anticodon loop

tRNA

Extending from the acceptor stem,


the 3' end of each tRNA has the
sequence CCA.

acceptor
stem

The appropriate
amino acid is
attached to the
ribose of the
terminal adenosine
(A, in red) at the 3'
end.
The anticodon
loop is at the

anticodon

Phe

tRNA

acceptor
stem

#46
(m7G)
#22
G

Tertiary base
Phe
pairs in tRNA

#13
C

#46
(m7G)
#22
G

#13
C

Tertiary base
Phe
pairs in tRNA

An example of non-standard H bond


interactions that help to stabilize the Lshaped tertiary structure of a tRNA, in ball &
stick & spacefill displays.
H atoms are not shown. (From NDB file 1TN2).

Some other RNAs,


including viral RNAs
& segments of
ribosomal RNA, fold
in pseudoknots,
tertiary structures
that mimic the 3D
structure of tRNA.
Pseudoknots are
similarly stabilized
by non-standard Hbond interactions.
Explore tRNAPhe
with Chime (PDB file

anticodon

tRNA

Phe

acceptor
stem

H
C

O
C

O
O

Amino acid

NH3+

H
C

CH2

ATP

Adenine

OH

H
OH

O
O

NH2

Aminoacyl-AMP

CH2

O
H

Adenine

OH

H
OH

PPi

Aminoacyl-tRNA Synthetases catalyze linkage of the


appropriate amino acid to each tRNA.
The reaction occurs in 2 steps.
In step 1, an O atom of the amino acid -carboxyl attacks
the P atom of the initial phosphate of ATP.

H
C

O
C

O
O

CH2

NH2

Aminoacyl-AMP

In step 2, the
2' or 3' OH of
the terminal
adenosine of
tRNA attacks
the amino acid
carbonyl C
atom.

OH

H
OH

tRNA
AMP

tRNA

Adenine

O
O

CH2

H
H

Adenine
H
H
OH

O
C

HC

NH3+

(terminal 3nucleotide
of appropriate tRNA)
Aminoacyl-tRNA

Aminoacyl-tRNA Synthetase - summary:


1. amino acid + ATP aminoacyl-AMP +
PPi
2. aminoacyl-AMP + tRNA aminoacyltRNA + AMP
The 2-step reaction is spontaneous overall,
because the concentration of PPi is kept low
by its hydrolysis, catalyzed by
Pyrophosphatase.

There is generally a different Aminoacyl-tRNA


Synthetase (aaRS) for each amino acid.
Accurate translation of the genetic code
depends on attachment of each amino acid to
an appropriate tRNA.
Each aaRS recognizes its particular amino
acid & tRNAs coding for that amino acid.
Identity elements: tRNA domains recognized
by an aaRS.
anticodon loop
Most identity elements are in the
acceptor stem & anticodon loop.
Aminoacyl-tRNA Synthetases arose
acceptor
early in evolution.
stem
tRNA
Early aaRSs probably recognized

tRNA
O
O

(terminal 3nucleotide
of appropriate tRNA)
CH2

There are 2 families


of Aminoacyl-tRNA
Synthetases:
Class I & Class II.

H
H

Adenine
H
H
OH

HC

NH3+

Aminoacyl-tRNA

Two different ancestral proteins evolved into


the 2 classes of aaRS enzymes, which differ in
the architecture of their active site domains.
They bind to opposite sides of the tRNA
acceptor stem, aminoacylating a different OH

Class I aaRSs:
Identity elements usually include residues of
the anticodon loop & acceptor stem.
Class I aaRSs aminoacylate the 2'-OH of
adenosine at their 3' end.
Class II aaRSs:
Identity elements for some Class II enzymes
do not include the anticodon domain.
Class II aaRSs tend to aminoacylate the 3'OH of adenosine at their 3' end.

Proofreading/quality control:
Some Aminoacyl-tRNA Synthetases are known to
have separate catalytic sites that release by
hydrolysis inappropriate amino acids that are
misacylated or mis-transferred to tRNA.
E.g., the aa-tRNA Synthetase for isoleucine (IleRS) a
small percentage of the time activates the closely
related amino acid valine to valine-AMP.
After valine is transferred to tRNAIle, to form ValtRNAIle, it is removed by hydrolysis at a separate
active site of IleRS that accommodates Val but not
the larger Ile.
In some bacteria, editing of some misacylated tRNAs
is carried out by separate proteins that may be
evolutionary precursors to editing domains of aatRNA Synthetases.

Some amino acids are modified after being


linked to a tRNA. Examples:
In prokaryotes the initiator tRNAfMet is first
charged with methionine.
Methionyl-tRNA formyltransferase then
catalyzes formylation of the methionine,
using tetrahydrofolate as formyl donor, to
yield formylmethionyl-tRNAfMet.
In some prokaryotes, a non-discriminating
aaRS loads aspartate onto tRNAAsn.
The aspartate moiety is then converted by
an amido-transferase to asparagine,
yielding Asn-tRNAAsn.
Glu-tRNAGln is similarly formed and

Some proteins contain


the unusual amino acid
selenocysteine (Sec),
with selenium substituting
for the S atom in cysteine.
There is a selenocysteine tRNA that differs
from other tRNAs, e.g., in having a slightly
longer acceptor stem & a unique modified base
in the anticodon loop.
tRNASec is loaded with serine via Seryl-tRNA
Synthetase.
The serine moiety is then converted to
selenocysteine by another enzyme, in a
reaction involving selenophosphate.

Other roles of aminoacyl-tRNA


synthetases:
In some organisms, Aminoacyl-tRNA
Synthetases (aaRSs) have evolved to
take on signaling roles in addition to the
catalytic role of joining an amino acid to
the correct tRNA.
Examples have been identified of
particular aaRSs that regulate
transcription, translation or intron
splicing through binding to DNA or RNA.

Proteolytic cleavage of the human


aaRSTyr yields a cytokine that stimulates
angiogenesis.
A truncated form of the human aaRSTrp
inhibits angiogenesis.
Regulation of apoptosis by the human
aaRSGln is dependent on the
concentration of its substrate glutamine.
Several mammalian Aminoacyl-tRNA
Synthetases associate with other proteins
to form large macromolecular
complexes whose roles are actively being
investigated.

Structure of tRNAs. (a) The primary structure of yeast alanine tRNA (tRNAAla), the first such sequence determined. This
molecule is synthesized from the nucleotides A, C, G, and U, but some of the nucleotides, shown in red, are modified
after synthesis: D = dihydrouridine, I = inosine, T = thymine, Y = pseudouridine, and m = methyl group. Although the exact
sequence varies among tRNAs, they all fold into four base-paired stems and three loops. The partially unfolded molecule
is commonly depicted as a cloverleaf. Dihydrouridine is nearly always present in the D loop; likewise, thymidylate,
pseudouridylate, cytidylate, and guanylate are almost always present in the TYCG loop. The triplet at the tip of the
anticodon loop base-pairs with the corresponding codon in mRNA. Attachment of an amino acid to the acceptor arm
yields an aminoacyl-tRNA. (b) Computergenerated three-dimensional model of the generalized backbone of all tRNAs.
Note the L shape of the molecule

A tRNA molecule. In this series of diagrams, the same tRNA moleculein this case a tRNA specific for the amino acid
phenylalanine (Phe)is depicted in various ways. (A) The cloverleaf structure, a convention used to show the complementary
base-pairing (red lines) that creates the double-helical regions of the molecule. The anticodon is the sequence of three
nucleotides that base-pairs with a codon in mRNA. The amino acid matching the codon/anticodon pair is attached at the 3
end of the tRNA. tRNAs contain some unusual bases, which are produced by chemical modification after the tRNA has been
synthesized. For example, the bases denoted Y (for pseudouridine) and D (for dihydrouridine) are derived from uracil. (B and
C) Views of the actual L-shaped molecule, based on x-ray diffraction analysis. Although a particular tRNA, that for the amino
acid phenylalanine, is depicted, all other tRNAs have very similar structures. (D) The linear nucleotide sequence of the
molecule, color-coded to match A, B, and C.

Structure of a tRNA-splicing endonuclease docked to a precursor tRNA. The endonuclease (a foursubunit enzyme) removes the tRNA intron (blue). A second enzyme, a multifunctional tRNA ligase (not
shown), then joins the two tRNA halves together

Processing of an Escherichia coli pre-tRNA. The example shown results in synthesis of tRNAtyr. The tRNA sequence in the
primary transcript adopts its base-paired cloverleaf structure and two additional hairpin structures form, one on either side of the
tRNA. Processing begins with the cut by ribonuclease E or F forming a new 3 end just upstream of one of the hairpins.
Ribonuclease D, which is an exonuclease, trims seven nucleotides from this new 3 end and then pauses while ribonuclease P
makes a cut at the start of the cloverleaf, forming the 5 end of the mature mRNA. Ribonuclease D then removes two more
nucleotides, creating the 3 end of the mature molecule. With this tRNA the 3-terminal CCA sequence is present in the RNA and is
not removed by ribonuclease D. With some other tRNAs this sequence has to be completely or partly added by tRNA
nucleotidyltransferase. Abbreviation: RNase, ribonuclease

Processing of transfer RNAs (A) Transfer RNAs


are derived from pre-tRNAs, some of which contain
several individual tRNA molecules. Cleavage at the
5 end of the tRNA is catalyzed by the RNase P
ribozyme; cleavage at the 3 end is catalyzed by a
conventional protein RNase. A CCA terminus is
then added to the 3 end of many tRNAs in a
posttranscriptional processing step. Finally, some
bases are modified at characteristic positions in the
tRNA molecule. In this example, these modified
nucleosides include dihydrouridine (DHU),
methylguanosine (mG), inosine (I), ribothymidine
(T), and pseudouridine (). (B) Structure of
modified bases. Ribothymidine, dihydrouridine, and
pseudouridine are formed by modification of
uridines in tRNA. Inosine and methylguanosine are
formed by the modification of guanosines.

Processing of tyrosine pre-tRNA involves four types of changes. A 14-nucleotide intron (blue) in the
anticodon loop is removed by splicing. A 16-nucleotide sequence (green) at the 5 end is cleaved by RNase P. U
residues at the 3 end are replaced by the CCA sequence (red) found in all mature tRNAs. Numerous bases in
the stem-loops are converted to characteristic modified bases (yellow). Not all pre-tRNAs contain introns that are
spliced out during processing, but they all undergo the other types of changes shown here. D = dihydrouridine; Y
= pseudouridine.

Mechanism of splicing in pre-tRNA. First, the pre-tRNA is cleaved at two places, on each side of the intron, thereby excising the
intron. The cleavage mechanism generates a 2,3-cyclic phosphomonoester at the 3 end of the 5 exon. The multistep
reaction joining the two exons requires two nucleoside triphosphates: a GTP, which contributes the phosphate group (yellow) for the
3 5 linkage in the finished tRNA molecule; and an ATP, which forms an activated ligase-AMP intermediate. The 2phosphate on the 5 exon is removed in the final step.

The structure of the aminoacyl-tRNA linkage. The carboxyl end of the amino acid forms an ester bond to
ribose. Because the hydrolysis of this ester bond is associated with a large favorable change in free energy, an
amino acid held in this way is said to be activated. (A) Schematic drawing of the structure. The amino acid is
linked to the nucleotide at the 3 end of the tRNA (see Figure 6-52). (B) Actual structure corresponding to boxed
region in (A). These are two major classes of synthetase enzymes: one links the amino acid directly to the 3-OH
group of the ribose, and the other links it initially to the 2-OH group. In the latter case, a subsequent
transesterification reaction shifts the amino acid to the 3 position. As in Figure 6-56, the R-group indicates the
side chain of the amino acid.

Helix Stacking in tRNA.


The four helices of the secondary structure of tRNA ()
stack to form an L-shaped structure

Editing of Aminoacyl-tRNA. The flexible CCA arm of an aminoacyl-tRNA can move


the amino acid between the activation site and the editing site. If the amino acid fits
well into the editing site, the amino acid is removed by hydrolysis.

Amino acid activation. The two-step process in which an amino acid (with its side chain denoted by R) is
activated for protein synthesis by an aminoacyl-tRNA synthetase enzyme is shown. As indicated, the energy of
ATP hydrolysis is used to attach each amino acid to its tRNA molecule in a high-energy linkage. The amino acid
is first activated through the linkage of its carboxyl group directly to an AMP moiety, forming an adenylated amino
acid; the linkage of the AMP, normally an unfavorable reaction, is driven by the hydrolysis of the ATP molecule
that donates the AMP. Without leaving the synthetase enzyme, the AMP-linked carboxyl group on the amino acid
is then transferred to a hydroxyl group on the sugar at the 3 end of the tRNA molecule. This transfer joins the
amino acid by an activated ester linkage to the tRNA and forms the final aminoacyl-tRNA molecule.

This figure depicts the


general molecular
function of an aminoacyl
tRNA synthetase. First,
the enzyme binds the
amino acid and joins it to
a molecular of AMP
while cleaving two
phosphate groups from
a molecule of ATP. Next,
the enzyme binds the
aminoacyl portion of this
complex to an
appropriate tRNA
molecule while releasing
the AMP molecule.

Hydrolytic editing. (A) tRNA synthetases remove their own coupling errors through
hydrolytic editing of incorrectly attached amino acids. As described in the text, the
correct amino acid is rejected by the editing site. (B) The error-correction process
performed by DNA polymerase shows some similarities; however, it differs so far as
the removal process depends strongly on a mispairing with the template

Transcription-control elements in genes


transcribed by RNA polymerase I (a) and III
(b,c). Assembly of transcription-initiation
complexes on these genes begins with the
binding of specific general transcription
factors to the control elements

The promoters of 5S rRNA and tRNA genes are downstream of the transcrip-tion initiation site.
Transcription of the 5S rRNA gene is initiated by the binding of TFIIIA, followed by the binding of
TFIIIC, TFIIIB, and the polymerase. The tRNA promoters do not contain a binding site for TFIIIA, and
TFIIIA is not required for their transcription. Instead, TFIIIC initiates the transcription of tRNA genes
by binding to promoter sequences, followed by the association of TFIIIB and polymerase. The TATAbinding protein (TBP) is a subunit of TFIIIB .

Ribosome Composition (S = sedimentation coefficient)


Ribosome
Source
E. coli

Whole
Ribosome
70S

Small
Subunit
30S
16S RNA
21 proteins

Rat
cytoplasm

80S

40S
18S RNA
33 proteins

Large
Subunit
50S
23S & 5S
RNAs
31 proteins
60S
28S, 5.8S, &5S
RNAs
49 proteins

Eukaryotic cytoplasmic ribosomes are larger


and more complex than prokaryotic
ribosomes. Mitochondrial and chloroplast

5S rRNA
crown view
displayed as
ribbons & sticks.

PDB 1FFK

Structures of large & small subunits of


bacterial & eukaryotic ribosomes have been
determined, by X-ray crystallography & by
cryo-EM with image reconstruction.
Consistent with predicted base pairing, X-ray
crystal structures indicate that ribosomal

Structure of the E. coli


Ribosome
large subunit

tRNA

EF-G

small subunit

mRNA
location

The cutaway view at right shows positions of


tRNA (P, E sites) & mRNA (as orange beads).
EF-G will be discussed later. This figure was
provided by Joachim Frank, whose lab at the
Wadsworth Center carried out the cryo-EM
and 3D image reconstruction on which the

Small Ribosomal Subunit


In the translation complex, mRNA threads through a
tunnel in the small ribosomal subunit.
tRNA binding sites are in a cleft in the small subunit.
The 3' end of the 16S rRNA of the bacterial small
subunit is involved in mRNA binding.
The small ribosomal subunit is relatively flexible,
assuming different conformations.
E.g., the 30S subunit of a bacterial ribosome was
found to undergo specific conformational changes
when interacting with a translation initiation factor.

Small
ribosomal
subunit of a
thermophilic
bacterium:
rRNA in
monochrome;
proteins in
varied colors.

30S ribosomal subunit

spacefill display

PDB 1FJF

ribbons

The overall shape of the 30S ribosomal subunit is largely


determined by the rRNA. The rRNA mainly consists of
double helices (stems) connected by single-stranded loops.
The proteins generally have globular domains, as well as
long extensions that interact with rRNA and may stabilize
interactions between RNA helices.

Large ribosome subunit:


The interior of the large
subunit is mostly RNA.
Proteins are distributed
mainly on the surface.

PDB 1FFK

Large
Ribosome
Subunit

Some proteins have long


tails that extend into the
interior of the complex.
These tails, which are
highly basic, interact
with the negatively
charged RNA.

"Crown" view with RNAs blue, in


spacefill; proteins red, as backbone.

PDB 1FFK

The active site domain


for peptide bond
formation is essentially
devoid of protein.

Large
Ribosome
Subunit

The peptidyl transferase


function is attributed to
23S rRNA, making this
RNA a "ribozyme."
"Crown" view with RNAs blue, in
spacefill; proteins red, as backbone.

Protein
synthesis takes
place in a cavity
within the
ribosome,
between small &
large subunits.
Nascent
polypeptides
emerge through a
tunnel in the
large subunit.
The tunnel lumen
is lined with rRNA

PDB 1FFK

Large ribosome subunit.


Backbone display with RNAs blue. View
from bottom at tunnel exit.

Catalysis of protein synthesis and movement


of the ribosome relative to messenger RNA are
accompanied by changes in ribosome
conformation.
EM & X-ray crystallographic studies, carried
out in the presence & absence of initiation &
elongation factors as well as inhibitors of
protein synthesis, have revealed
conformational changes in rRNA.
Thus rRNA participates in conformational
coupling in addition to its structural &
catalytic roles.
tRNAs also undergo substantial
conformational changes within ribosomal

Pengertiaan splicingRNA
Splicing
RNA
merupakan
pemotongan
intron
untuk
menggabungkan exon dalam peristiwa
transkripsi. Sebelum membahas lebih
jauh tentang splicing, saya akan
mengulang penjelasan tentang intron
dan exon pada gen makhluk hidup.

Gen merupakan sandi genetik yang


menyimpan informasi yang menentukan sifat
tertentu dari makhluk hidup. Gen terdiri atas
bagian yang akan diekspresikan yang disebut
exon dan bagian yang tidak diekspresikan
yang disebut intron. Bagin exon nantinya akan
ditranskripsikan menjadi RNAm, sedangkan
intron terletak di antara exon, sehingga harus
dipotong agar tercipta RNAm yang runtut.

Bagaimana intron
dipotong?

Gen akan ditranskripsikan menjadi RNAm terlebih


dahulu untuk membentuk protein. Gen yang mengandung
intron akan ditranskripsikan menjadi pre-RNAm yang
harus menjalani splicing agar terbentuk RNAm matang.
Proses splicing membutuhkan bantuan dari RNSsn (RNA
small nucllear) dan protein-protein tertentu.

RNAsn dan kompleks protein akan bergabung


membentuk spliceosome yang akan memotong intron.
Spliceosome akan mengikat ujung 5' dan 3' dari intron,
membentuk lengkungan dan kemudian memotong intron
tersebut. Hilangnya intron akan membuat exon bersatu
sehingga pre-RNAm berubah menjadi RNAm yang siap
diranslasikan menjadi protein.

INISIASI
Tahap inisiasi terjadi karena adanya tiga komponen yaitu mRNA, sebuah tRNA
yang memuat asam amino pertama dari polipeptida, dan dua sub unit
ribosom.
mRNA yang keluar dari nukleus menuju sitoplasma didatangi oleh ribosom,
kemudian mRNA masuk ke dalam celah ribosom. Ketika mRNA masuk ke
ribosom, ribosom membaca kodon yang masuk. Pembacaan dilakukan
untuk setiap 3 urutan basa hingga selesai seluruhnya. Sebagai catatan
ribosom yang datang untuk mebaca kodon biasanya tidak hanya satu,
melainkan beberapa ribosom yang dikenal sebagai polisom membentuk
rangkaian mirip tusuk satu, di mana tusuknya adalah mRNA dan daging
adalah ribosomnya. Dengan demikian, proses pembacaan kodon dapat
berlangsung secara berurutan. Ketika kodon I terbaca ribosom (misal
kodonnya AUG), tRNA yang membawa antikodon UAC dan asam amino
metionin datang. tRNA masuk ke celah ribosom.
Ribosom di sini berfungsi untuk memudahkan perlekatan yang spesifik antara
antikodon tRNA dengan kodon mRNA selama sintesis protein. Sub unit
ribosom dibangun oleh protein-protein dan molekul-molekul RNA ribosomal.

Elogasi
Pada tahap elongasi dari translasi, asam amino-asam amino ditambahkan satu per
satu pada asam amino pertama (metionin). Ribosom terus bergeser agar mRNA
lebih masuk, guna membaca kodon II. Misalnya kodon II UCA, yang segera
diterjemahkan oleh tRNA berarti kodon AGU sambil membawa asam amino serine.
Di dalam ribosom, metionin yang pertama kali masuk dirangkaikan dengan serine
membentuk dipeptida.
Ribosom terus bergeser, membaca kodon III. Misalkan kodon III GAG, segera
diterjemahkan oleh antikodon CUC sambil membawa asam amino glisin. tRNA
tersebut masuk ke ribosom. Asam amino glisin dirangkaikan dengan dipeptida
yang telah terbentuk sehingga membentuk tripeptida. Demikian seterusnya proses
pembacaan kode genetika itu berlangsung di dalam ribobom, yang diterjemahkan
ke dalam bentuk asam amino guna dirangkai menjadi polipeptida.
Kodon mRNA pada ribosom membentuk ikatan hidrogen dengan antikodon molekul
tRNA yang baru masuk yang membawa asam amino yang tepat. Molekul mRNA
yang telah melepaskan asam amino akan kembali ke sitoplasma untuk mengulangi
kembali pengangkutan asam amino. Molekul rRNA dari sub unit ribosom besar
berfungsi sebagai enzim, yaitu mengkatalisis pembentukan ikatan peptida yang
menggabungkan polipeptida yang memanjang ke asam amino yang baru tiba.

Terminasi

Tahap akhir translasi adalah


terminasi. Elongasi berlanjut hingga
kodon stop mencapai ribosom. Triplet
basa kodon stop adalah UAA, UAG,
dan
UGA.
Kodon
stop
tidak
mengkode
suatu
asam
amino
melainkan bertindak sinyal untuk
menghentikan translasi. Polipeptida
yang dibentuk kemudian diproses
menjadi protein.

Perbedaan sintesis protein eukariot


dan prokariot

Kontrol Positif dan Negatif


Kontrol Negatif(protein dibutuhkan untuk memblock
transkripsi)
a.Operon dinonaktifkan oleh produk ekspresi gen
regulator(represor)
b.Repsesor melekat pada operatortranskripsi
dihambat
Kontrol Positif(membutuhkan suatu protein untuk terjadinya
transkripsi)
a.Operon diaktifkan oleh produk ekspresi gen
regulator(aktivator)
b.Aktivator melekat pada operatortranskripsi
berjalan

Promotor adalah urutan DNA spesifik yang


berperan dalam mengendalaikan
transkripsi gen struktural dan terletak di
daerahupstream(hulu) dari bagian
struktural gen.
Operator merupakan urutan nukelotida
yang terletak di antara promotor dan
bagian struktural dan merupakan tempat
pelekatan protein represor (penekan atau
penghambat ekspresi gen).

Ekspresi gen dan


mekanisme
pengendaliannya
A. Operon dan regulon
B. Pengendalian pasca transkripsi
C. Pengendalian pasca translasi dan
modifikasi protein
D. Pengendalian global dan sinyal
transduksi
67

Dari Gen ke Protein

68

Ekspresi Gen

69

Animasi Translasi

70

Regulasi Gen pada Bakteri


Bakteri mempunyai beribu-ribu gen
Tidak semua ditranskripsi pada waktu
yang sama
Bila itu dilakukan maka akan
membuang energi yang banyak
Namun beberapa gen ditranskripsi
sepanjang waktu Siapa dia ?
housekeeping genes

Gen lain diekspresikan sebagai


tanggapan (respon) akibat terjadinya
perubahan lingkungan
71

Regulasi Gen pada Bakteri


Regulasi transkripsi
Jika suatu protein (yang dikodekan
oleh gen) diperlukan, maka gen akan
ditranskripsi
Jika suatu protein (yang dikodekan
oleh gen) Tidak diperlukan, maka gen
akan Tidak akan ditranskripsi

72

Unit Transkripsi pada


Bakteri
Operon

promoter
operator
-35
-10

TTGACA
TATAAT (Pribnow box)
73

Regulasi pada Transkripsi


Kontrol Positif atau Negatif
Positif - membutuhkan suatu protein
untuk terjadinya transkripsi
Negatif - protein dibutuhkan untuk
mem block transkripsi

Model operon pada bakteri


Lac operon
74

Regulasi Lac operon


Merupakan Kontrol Negatif
Repressor protein (lac I gene product)
Berikatan pada operator region dari
Lac operon
Mencegah terjadinya proses transkripsi

75

Induksi repressed operon

(allolactose)

(cant bind)

(lac repressor)

o
76

Regulasi Lac operon


Repressor berikatan pada operator
TIDAK terjadi transkripsi

Effector berikatan pada Repressor


Repressor tidak dapat berikatan dengan
operator
Terjadi Transkripsi

Berkurangnya konsentrasi Effector


Repressor akan bebas untuk berikatan
dengan operator
TIDAK terjadi transkripsi
77

Regulasi Lac operon


Satu gen yang berekspresi dari lac
operon
-galactosidase
Memecah laktosa (dan allolactose)
menjadi glukosa dan laktosa
LACTOSE
(allolactose)
(effector)

-gal

Glucose
&
Galactose
78

Repressi Operon Bakteri

Repressor

complex

co-repressor (tryptophan)
Biosynthetic
Operons
(Trp Operon)

79

Aktivasi & Deaktivasi Operon

effector
Activator

inactive
complex

80

Regulasi Transkripsi Eukariotik


Beberapa gen ditranskripsi pada hampir
semua sel
housekeeping genes

Sifat unik dari sel itu disebabkan oleh


ekspresi gen-gen spesifik yang
terkandung dalam sel tersebut
cell-specific expression
tissue-specific expression
81

Regulasi Transkripsi Eukariotik


Kemampuan untuk aktivasi dan
penekanan gen menjadi bagian
yang esensial untuk memelihara
kespesifikan sel
neuron

hepatocyte

skin
82

Regulasi Transkripsi Eukariotik


Lebih kompleks dibanding bakteri
Pengendalian dimediasi oleh proteinprotein yang diklasifikasikan sebagai
Transcription Factors - TF :
Basal TF - diperlukan oleh semua gen
Specific TF - menentukan spesifitas
ekspresi
Activator - meningkatkan ekspresi
Repressor - menurunkan ekspresi
83

Skema Promoter Gen Eukariot


SP1

TFIID

Hampir semua mempunyai TATA box pada -25


Posisi TF binding sites lebih upstream
GC box CCAAT box etc.

84

Penerapan
Merekayasa promoter yang
berbeda di depan coding
sequences yang diinginkan, dapat
menghasilkan:
a. Regulasi yang berbeda
b. Meningkatkan laju ekspresi
c. Mengubah waktu ekspresi
d. Ekspresi terjadi di jaringan yang

berbeda
e. Ekspresi pada organisme yang
85
berbeda

Animasi Regulasi

Operon Lac

Eukariotik Control Expr.

Operon
86

Signal Transduksi

87

signal transduction refers to any process by which a cell converts one kind of
signal or stimulus into another, most often involving ordered sequences of
biochemical reactions inside the cell, that are carried out by enzymes, activated
by second messengers resulting in what is thought of as a "signal trandusction
pathway". Such processes are usually rapid, lasting on the order of milliseconds
in the case of ion flux, minutes for the activation of protein and lipid mediated
kinase cascades, or hours and days in terms of gene expression. In many signal
transduction processes, the number of proteins and other molecules participating
in these events increases as the process emanates from the initial stimulus,
resulting in a "signal cascade" and often results in a relatively small stimulus
eliciting a large response. This is referred to as amplification of the signal.
In bacteria and other single-cell organisms, the variety of a signal transduction a
processes of which the cell is capable influences how many ways it can react and
respond to its environment. In multicellular organisms , a multitude of different
signal transduction processes are required for coordinating the behavior of
individual cells to support the function of the organism as a whole. As may be
expected, the more complex the organism, the more complex the repertoire of
signal transduction processes the organism must possess. Thus, sensing of both
the external and internal environment at the cellular level, relies on signal
transduction. Many disease processes such as diabetes, heart disease,
autoimmunity and cancer arise from defects in signal transduction pathways,
further highlighting the critical importance of signal transduction to biology as
well as medicine.

88

Anda mungkin juga menyukai