Gene Expression

1
EKSPRESI GEN
Merupakan proses pengungkapan bahan informasi
genetik yang tersusun sebagai gen ke dalam bentuk
protein (merupakan proses transkripsi gen tertentu
yang diikuti proses translasi)

DNA --> transcription --> RNA -- translation -->

Protein
 Gene Structure
Eukaryotic gene structure: Most
eukaryotic genes in contrast to
typical bacterial genes, the coding
sequences (exons) are interrupted
by noncoding DNA (introns).
The gene must have (Exon; start
signals; stop signals; regulatory
control elements).

 The average gene 7-10 exons

spread over 10-16kb of DNA.

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 Protein Synthesis: Four stages
 Transcription
 RNA processing
 Translation
 Post-translation processing
Transcription
 Synthesis of an RNA
that is complementary to
one of the strands of
DNA.
Translation
 Ribosomes read a
messenger RNA and
make protein according
to its instruction.
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INISIASI
ELONGASI
TERMINASI
 TRANSKRIPSI PADA PROKARIOT

•Tidak memerlukan TATA Box.

(RNA Polimerase dapat
langsung menempel pada
promotor dan memulai
transkripsi)
•Pada Prokariot awal Inisiasi
RNA Polimerase dibantu oleh
faktor sigma
•Transkripsi dimulai dengan
adanya binding dari RNA
polimerase ke promotor dalam
Proses transkripsi langsung
menghasilkan mRNA (tanpa
adanya splicing)
TRANSKRIPSI PADA EUKARIOT

 Terjadi di dalam nukleus yang

kemudian ditranslasi di
sitoplasma
 Memerlukan TATA Box dan
faktor-faktor Transkripsi sebagai
tempat menempelnya RNA
Polimerase dan memulai
Transkripsi
 Melibatkan 3 macam RNA
Polymerase (I, II, dan III)
 Transkripsi menghasilkan “pre-
mRNA” yang kemudian
Mengalami “splicing” untuk
menjadi mRNA
 Pada organisme prokariot mempunyai 1 tipe RNA
polimerase
 Pada eukariotik memiliki 3 tipe RNA polimerase :
1. RNA polimerase I : lokasi ada di nukleolus,
mengkatalisis sintesis r RNA
2. RNA polimerase II : lokasi di nukleoplasma,
mengkatalisis sintesis hn RNA atau pre-m RNA
3. RNA polimerase III : lokasi di nukleoplasma,
mengkatalisis sintesis sn RNA dan t RNA
 RNA Processing (Pre-mRNA → mRNA)
 Capping
 Splicing
 Addition of poly A tail
 Addition of poly A tail:
Synthesis of the poly (A) tail
involves cleavage of its 3' end
and then the addition of about 40-
200 adenine residues to form a
poly (A) tail.
 Capping
 The cap structure is added to
the 5' of the newly transcribed
mRNA precursor in the nucleus
prior to processing and
subsequent transport of the
mRNA molecule to the
cytoplasm.
 Splicing:
 Step by step removal of pre
mRNA and joining of
remaining exons; it takes place
on a special structure called
spliceosomes.
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 Eukaryotic RNA is processed
before leaving the nucleus
 Noncoding DNA
Exon Intron Exon Intron Exon

segments, introns, Transcription

Cap Addition of cap and tail
are spliced out
RNA
 A cap and a tail are transcript
with cap Introns removed Tail
and tail
added to the ends
Exons spliced together

mRNA

Coding sequence
NUCLEUS

CYTOPLASM

Figure 10.10
 Alternative Splicing
Alternative splicing:
is a very common
phenomenon in higher
eukaryotes. It is a way to
get more than one
protein product out of
the same gene and a way
to control gene
expression in cells.

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 Translation of nucleic acids into amino acids

 The “words” of the DNA “language” are triplets of bases

called codons
 The codons in a gene specify the amino acid sequence of a
polypeptide
 Gene 1 Gene 3

DNA molecule

Gene 2

DNA strand

TRANSCRIPTION

RNA

Codon
TRANSLATION

Polypeptide
Amino acid
Figure 10.7
Virtually all organisms share the same genetic
code “unity of life” Second Base
U C A G
UUU UCU UAU UGU U
phe tyr cys
UUC UCC UAC UGC C
U ser
UUA UCA UAA stop UGA stop A
leu
UUG UCG UAG stop UGG trp G
CUU CCU CAU his CGU U
CUC CCC CAC CGC C
C leu pro arg

Third Base
First Base

CUA CCA CAA gln CGA A

CUG CCG CAG CGG G
AUU ACU AAU AGU U
asn ser
AUC ile ACC AAC AGC C
A thr
AUA ACA AAA AGA A
lys arg
AUG met (start) ACG AAG AGG G
GUU GCU GAU GGU U
asp
GUC GCC GAC GGC C
G val ala gly
GUA GCA GAA GGA A
glu
GUG GCG GAG GGG G
 An exercise in translating the genetic code

Transcribed strand

DNA

Transcription

RNA

Start Stop
codon Translation codon

Polypeptide
Figure 10.8B
 Transfer RNA molecules serve as interpreters during
translation
Amino acid attachment site
 In the cytoplasm, a
ribosome attaches to
the mRNA and
translates its message
into a polypeptide
 The process is aided Hydrogen bond

(dibantu) by transfer
RNAs
RNA polynucleotide chain

Anticodon
Figure 10.11A
 Each tRNA molecule has a triplet anticodon on one end
and an amino acid attachment site on the other

Amino acid
attachment
site

Anticodon
Figure 10.11B, C
 Ribosomes build polypeptides

Next amino acid

Growing to be added to
polypeptide polypeptide
tRNA P site A site
molecules
Growing
Large polypeptide
subunit
tRNA

P A
mRNA
mRNA
binding
site
Codons

mRNA Small
subunit

Figure 10.12A-C
 An initiation codon marks the start of
an mRNA message

AUG = methionine

Start of genetic message

End

Figure 10.13A
 mRNA, a specific tRNA, and the ribosome subunits
assemble during initiation

Large
Initiator tRNA ribosomal
subunit
P site
A site

Start
codon Small ribosomal
mRNA subunit

1 2

Figure 10.13B
 Elongation
 The mRNA moves a codon at a time relative to the
ribosome
 A tRNA pairs with each codon, adding an amino acid to
the growing polypeptide
 A STOP codon causes the mRNA-ribosome complex to fall
apart
 Amino acid

Polypeptide
A
P site site
Anticodon

mRNA
1 Codon recognition

mRNA
movement

Stop
codon

New 2 Peptide bond

peptide formation
bond

3 Translocation Figure 10.14

Table 14.2
Types of RNA

Type of RNA Functions in Function

Messenger RNA Nucleus, Carries DNA
(mRNA) migrates sequence
to ribosomes information to
in cytoplasm ribosomes

Transfer RNA Cytoplasm Provides linkage

(tRNA) between mRNA
and amino acids;
transfers amino
acids to ribosomes

Ribosomal RNA Cytoplasm Structural

(rRNA) component
of ribosomes
 New
peptide
Growing bond Stage 4 Elongation
polypeptide forming
A succession of tRNAs
add their amino acids to
the polypeptide chain as
the mRNA is moved
through the ribosome,
one codon at a time.
Codons
mRNA
Polypeptide

Stage 5 Termination
The ribosome recognizes
a stop codon. The poly-
peptide is terminated and
Stop Codon released.

Figure 10.15 (continued)

 The Genetic Code

 The sequence of codons in

the mRNA defines the
primary structure of the
final protein.

 Three nucleotides in
mRNA (a codon)specify
one amino acid in a
protein.

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 The Genetic Code
 The triplet sequence of
mRNA that specify
certain amino acid.
 64 different combination of
bases; 61 of them code for
20 amino acids (AA); the
last three codon
(UAG,UGA,UAA) don not
code for amino acids; they
are termination codons.
 Degenerate
 More than on triplet codon
specify the same amino
acid.
 Unambiguous
 Each codon specifies a
particular amino acid, the
codon ACG codes for the
amino acid threonine, and
only threonine.
 Non overlapping
 This means that
successive triplets are
read in order. Each
nucleotide is part of only
one triplet codon.
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DNA Codon RNA Codon

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 Translation

Translation is the process by

which ribosomes read the
genetic message in the mRNA
and produce a protein product
according to the message's
instruction.

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Pengemasan dan Prosesing
 Rantai polipeptida mulai menggulung, melipat secara
spontan membentuk protein fungsional suatu
konformasi yg spesifik

 Protein menjadi berfungsi melalui proses post

translasi dengan cara dimodifikasi, misal dengan
penambahan gugus prostetik, reaksi karboksilasi,
fosforilasi, penghilangan urutan penanda dll
 MODIFIKASI PASCA TRANSLASI
• Modifikasi terminal amino dan terminal karboksil
gugus fornil, residu metionin pemula dan 1 atau lebih
residu berikutnya sering tidak muncul pada produk
protein (residu terminal amino/karboksil mengalami
metilasi)
• Terlepasnya urutan memberi isyarat
urutan pengisyarat (untuk mengarahkan protein ke tujuan
akhir) dilepaskan oleh peptidase spesifik
• Fosforitasi aa hitrosi
fosforritasi gugus hidrosi residu serin untuk mengatifkan
enzim-enzim tertentu
 Reaksi karboksilasi
penambahan gugus karboksil pada aa aspartat dan
glutamat
 Metilasi gugus R
residu lisin dan gutamate mengalami metilasi
 Pengikatan rantai sisi karbohitrat
rantai sisi karbohirat gliprotein diikat secara kovalen selama
atau setelah sintesis protein