TRANSKRIPSI

SINTESIS RNA : mRNA, tRNA, rRNA

Dyah Ayu Oktavianie

PKH-UB

Study objectives
You will need to know ALL the concepts and details in this lecture.
1. What are the three main types of RNA and what are their functions?
2. Understand how the following terms apply to RNA structure: phosphodiester
bonds, 5' and 3 ends, nucleosides, complementary base pairing, stem loops.
3. Compare and contrast DNA and RNA structure.
4. What is a gene? What is gene expression? *Understand transcription,
translation, and RNA processing in both prokaryotes and eukaryotes.
5. Define operons and polycistronic messages. How do they function in
prokaryotic gene expression?
6. *Compare and contrast the features of prokaryotic and eukaryotic gene
expression. Do eukaryotes have operons? What are exons, introns, primary
transcripts, capping, tailing, and splicing. What is the 5' cap (methylguanosine
cap)? How and when is the poly-A tail added to the transcript? Where does
eukaryotic RNA processing occur?
7. Understand the structure and function of promoters and terminators in
transcription. Contrast intrinsic terminators and rho-dependent terminators.
8. Know the subunit structure of bacterial RNA polymerase and the sigma cycle.

DOGMA SENTRAL

DNA PEMBAWA INFORMASI

GENETIK
TRANSKRIPSI (SINTESIS mRNA, tRNA,
rRNA)
TRANSLASI (SINTESIS PROTEIN)
PENGENDALIAN

TRANSKRIPSI
Awal sintesis RNA, perpanjangan dan
terminasi
r- RNA
80%
t- RNA
15%
m- RNA 5%

A. Functions of the major RNAs

1. messenger RNAs (mRNA) contain

genetic information to encode a protein
phe

2. transfer RNAs (tRNA) act as adapters

between the mRNA nucleotide code and
amino acids during protein synthesis
3. ribosomal RNAs (rRNA) are structural and
catalytic component of ribosomes

2. The phosphodiester bonds

of RNA are analogous to those
of DNA.

3. The 5' and 3' ends

of RNA are analogous to
those of DNA.

TB

P-O-C

5' end

ring numbering
system for ribose
5'
-C O
1
4

phosphodiester
bond
O
O-

3 end

OH
O
P

3
O

HO

O
OH

RNA

TB

PERBEDAAN STRUKTUR DNA - RNA

KOMPOSISI BASA DNA & RNA

DNA

A
T
G
C

RNA
25%
33
24
18

U
A
C
G

25 %
32
23
20

Complementary base pairing

CCCTTTGGGAAA

DNA

GGGAAACCCUUU

RNA

GGGAAACCCUUU

RNA

CCCUUUGGGAAA

RNA

hydrogen
bonding

TB

Gene numbers
group
viruses
prokaryotes
eukaryotes

approximate
gene number
4-200
500-12,000
5,000-125,000
TB

Any given species has a unique set

of genes that confers a unique set
of properties.
Proteins and RNAs determine all of the
characteristics of organisms and cells.
Example: Escherichia coli has 4405 genes
~117 encode RNAs (tRNA, rRNA)
~4288 encode proteins

TB

Gene expression in prokaryotes

1. Expression of single genes

Ex.1: a single gene that encodes a protein

1 gene
transcription
translation

1 mRNA

1 polypeptide

TB

Ex. 2: a single gene that encodes one

rRNA or tRNA

1 gene
transcription

1 RNA
RNA processing

degraded

1 tRNA etc.
TB

2. Expression of operons
operon
two or more genes
transcribed together

C
DNA

transcription

polycistronic message
a single RNA molecule that
represents more than one gene

polycistronic
mRNA

TB

b. Operons can encode several

rRNA molecules.
1 operon
1 polycistronic
RNA
processing

rRNA

rRNA
degraded

2 or more rRNAs TB

Important points

The details of organization,

processing and degradation are
different for different RNAs.

Most prokaryotes use operons.

Operons are used to coordinate
gene expression and often contain
genes of related function.
TB

Prokaryotic transcription
A. overview
B. transcribed regions
C. RNA polymerase
D. promoters
E. terminators
F. sigma factor
TB

Overview of prokaryotic transcription

RNA synthesis from a DNA template
typical
gene dsDNA

RNA polymerase

primary transcript complementary to

one strand of the coding region

TB

Defined regions are transcribed

upstream transcribed downstream
region
region
region
gene dsDNA
promoter
(RNA polymerase
binding site)

transcription
start site

termination
site

TB

GENA

Promoter
Operator
Lokus penempelan aktivator
Struktural gena : mRNA
Terminator

Polisistronik

Monosistronik

TRANSKRIPSI = SINTESIS RNA

(mRNA, tRNA, rRNA)
Polimerase RNA
ATP, CTP, GTP, UTP

Urutan pada DNA:

Promoter
Operator
Situs penempelen aktivator
Terminator

Gambaran dasar sintesis RNA

Prekursor : ATP CTP GTP UTP

Polimerase RNA
Reaksi polimerisasi
Urutan RNA ditentukan DNA
Salah satu untai DNA sebagai catakan
Untai RNA tumbuh dari 3
Polimerase mulai sintesis tanpa primer

C. RNA polymerase is the enzyme that

synthesizes RNA from a DNA template.

RNA polymerase

gene,or
operon

DNA template
complementary RNA
TB

+
+

completed
transcript

TB

Subunit structure of bacterial

RNA polymerase
core enzyme
'

holoenzyme
'

The holoenzyme includes one

of several sigma factors.

TB

Binding of polymerase to Template

DNA
Polymerase binds nonspecifically to DNA with
low affinity and migrates, looking for promoter.
Sigma subunit recognizes promoter sequence.
RNA polymerase holoenzyme and promoter
form "closed promoter complex" (DNA not
unwound).
Polymerase unwinds about 12 base pairs to
form "open promoter complex.

The sigma factor cycle

Sigma factors ( ) are a subunit
of RNA polymerase.
Sigma factors are needed for promoter
binding, but after transcription starts they
dissociate.

TB

INISIASI

RNA pol holoenzyme (core + sigma)

sigma
factor

core enzyme
sigma factor
RNA (~10 nucleotides)

TB

termination
RNA

core enzyme
sigma
holoenzyme

TB

Upstream region of the lactose operon

AATGTGAGTTAGCTCACTCATTA
-35 region

GCACCCCAGGCTTGACATTTATG
-10 region (Pribnow)

TTCCGGCTCGTATGTTGTGTGGA

ATTGTGAGCGGATAACAATTTCA
Shine-dalgarno (RBS)

ACAGGAAAGAGCTATGACC...
Translation start site

Transcription
start site

romoters
es on DNA where RNA
lymerase binds to start transcription
upstream transcribed downstream
region
region
region
gene dsDNA

promoter

transcription
start site

termination
site

TB

Typical bacterial 70 promoter

TTGACA
AACTGT

TATAAT
ATATTA

TTGACA =
-35 consensus
sequence

TATAAT =
-10 consensus
sequence
*also called Pribnow box;
~ 10 bases before start
site of transcription TB

PROMOTER PROKARIOT

PROMOTER PROKARIOT & EUKARIOT

PERPANJANGAN TRANSKRIPSI

Transcriptional terminators

DNA region that mediates the

termination of transcription.
termination
site
gene dsDNA

region where
terminators are
usually found

TB

1. Intrinsic terminator
DNA encoding an RNA that forms
a stem loop followed by a run of "U"s
that is used for transcriptional termination.

RNA

UUUU
3' end of RNA

TB

Intrinsic terminator function

The RNA stem loop

binds to RNA pol and
causes termination
Important fact: Intrinsic terminators must be
transcribed in order to function.

TB

2. Rho-dependent terminator
A DNA site where RNA polymerase
pauses and transcription
is terminated by Rho protein

TB

Rho protein binds RNA then moves

along RNA until it contacts RNA pol and
terminates transcription

Rho protein

RNA pol
pauses at
Rho
termination site
TB

Heksamer Rho
Rho menempel RNA bergerak pada untai
tunggal mRNA dengan menghidrolisa ATP
Ketika Rho mencapai polimerase,
memecah hibrid RNA-DNA

tRNA assists in translation

Transfer RNA takes a specific amino acid to the
ribosome
Single stranded but has areas where bases hydrogen
bond to each other
Cloverleaf shape
3 end picks up the amino acid
Which amino acid is determined by 3 bases opposite the
amino acid called an anti-codon
Anti-codons pair with complementary codons in mRNA at
a ribosome

(a) Structure of 16S rRNA

(b) Structure of 23 S rRNA

STABILITAS RNA
mRNA t 2-3 menit,
cara pengendalian