Dasar-dasar Ekspresi Gen

GEN

sekuen DNA dengan panjang minimum tertentu yang mengkode urutan
lengkap asam amino suatu polipeptida, atau RNA (mRNA, tRNA, rRNA)

Awal transkripsi
ATG

Promoter

STOP

Gen Struktural
Struktur dasar gen

Terminator

A Eukaryotic Gene
How are eukaryotic genes different from prokaryotic genes?

Overview

RNA is transcribed, using the rules of complementary
base pairing, from the template strand of DNA.

Most genes code for proteins; a smaller fraction
encode untranslated functional RNAs.

The nucleotide sequence of the gene determines the
order of amino acids in a protein, which determines
shape, size, and protein function.

mRNA is translated in groups of three nucleotides
(codon) at the ribosome through pairing of tRNA
anticodon with the mRNA codon.

Replikasi DNA

Replikasi RNA

Transkripsi

DNA

RNA

Transkripsi balik
(Reverse transcription)

Dogma Biologi Modern

(Central Dogma)

Transla
i

PROTEIN


galaktosidase

Transfer of Information

DNA  RNA  polypeptide {Central Dogma}

originally believed this transfer of information was
in one direction only

RNA can go back to DNA via reverse
transcription; no transfer from protein to RNA

Complementary base pairing transfers
information

during transcription to form RNA

during translation to form protein

The importance of weak bonds in information
transfer

RNA

1st step in process of information transfer is
transcription

Transcription: copying nucleotide sequence of DNA
into RNA

forms RNA transcript

DNA may be transcribed multiple times

RNA

single-stranded polynucleotide

contains ribose sugar

contains the pyrimidine uracil (U)

hydrogen bonds with A

5 and 3 ends critically important

Classes of RNA

Informational RNA: protein encoding mRNA

primary transcript in prokaryotes

processed transcript in eukaryotes

5 and 3 end modification

intron removal

translated into amino acid sequence

Functional (structural) RNA

tRNA: transports amino acid to ribosome; Cricks adaptor

rRNA: structural and catalytic component of ribosomes

snRNA: structural and catalytic component of spliceosome
snRNPs

snoRNA: small nucleolar RNA involved in maturation of
rRNA

scRNA: directs protein traffic in cytoplasm

Transcription

RNA polymerase catalyzes RNA synthesis

uses one DNA strand as template

always the same strand for a given gene

locally unwinds DNA

adds free nucleotides to growing RNA strand at 3 end

5 to 3 RNA synthesis

template read 3 to 5

uses rules of base pairing to synthesize complementary
RNA molecule

starts RNA chain de novo

Transcript is identical in sequence to nontemplate

strand, except Ts replaced by Us

Transcription is asymmetric only one strand of the DNA is

transcribed into RNA; the template strand
The RNA transcript has the same sequence as the
nontemplate strand

RNA is synthesized in a 5 to 3 direction only

The template strand is read in the 3 to 5 direction

Either strand of the DNA can be used as the template strand

for transcription
However, in any one gene only one strand of the DNA serves as
the template for transcription

DNA

RNA polymerases

Prokaryotes: single RNA polymerase

Transcribes mRNA, rRNA and tRNA

Transcription and translation are coupled

Eukaryotes: three RNA polymerases

RNA polymerase I transcribes rRNA genes

RNA polymerase II transcribes protein-encoding genes; i.e.
makes mRNA

primary transcript will be processed

RNA polymerase III transcribes tRNA genes and 5S rRNA
genes

Transcription and translation occur in separate

compartments of the eukaryotic cell

In organelles they occur in the same compartment

Transcription steps

Initiation

at 5 end of gene

binding of RNA polymerase to promoter

unwinding of DNA

Elongation

addition of nucleotides to 3 end of growing chain

governed by rules of complementary base pairing

energy from NTP substrates

Termination
5UTR
3UTR
coding
region

at 3 end of gene

terminator loop (prokaryote) or processing enzyme

Translation

mRNA is translated at the ribosome using tRNA as an
adaptor molecule

nucleotide sequence is read three nucleotides at a
time

each triplet is called a codon

each amino acid has one or more codons

64 possible codons (4 4 4) = genetic code

used by all organisms with few exceptions

no punctuation except start and stop

Genetic code specifies 20 different amino acids
(sometimes selenocysteine)

Translation



 

What are some features
of the genetic code?
Each codon specifies a
single amino acid.
The code is degenerate
since there are multiple
codons for each amino
acid with 2 exceptions.
The multiple codons for
each amino acid begin
with the same two
nucleotides.

fMet

mRNA

AUG

50S subunit, H2 0, GTP

GDP, P i

fMet

mRNA

AUG

Pembentukan komplek inisiasi 70S

A
Penyisipan aminoasil-tRNA
oleh EF-Tu

fMet

AUG CGA

GCU

mRNA

fMet Arg

AUG CGA
GTP

GCU

mRNA

GDP + P i

fMet
Arg

fMet
Translokasi dibantu
oleh EF-G

AUG CGA GCU

Arg

mRNA
GTP

AUG CGA

GDP + P i

Proses pemanjangan rantai polipeptida selama sintesis protein

GCU

mRNA