Expression of Genetic Information : From

Transcription to Translation
Prof. DR. dr. Hadyanto Lim, M.Kes, SpFK, FESC, FIBA
Department of Pharmacology and Molecular Biology
Faculty of Medicine, Methodist University of Indonesia - Medan
Molecular Biology Research, Postgraduate School,
University of Sumatra Utara - Medan

The DNA Double Helix
The Flow of Information in a Eukaryotic Cell
DNA RNA Protein
The pathway from DNA to Protein


How is the relationship
between genes and proteins ?
A Scottish Physician
1908 Archibald Garrot, reported the
symptoms exhibited by persons with certain
rare inherited diseases were caused by the
absence of specific enzyme. The disease was
alcaptonuria, because the urine becomes
dark on exposure to air.

Garrod found that persons with alcaptonuria
lacked an enzyme in their blood.

Lack of homogentisate oxidase in alcaptonuria
Inborn error of metabolism by Garrod
Relationship between a genetic defect, a specific
enzyme, and a specific metabolism condition.

Hypothesis
One gene - one enzyme.
One gene - one polypeptide.
Modification
A single gene often generates a variety of
polypeptide.
The Beadle Tatum Experiment in 1940s
Conclusion of the Beadle Tatum Experiment
The cell have an enzymatic deficiency that prevents them
from synthesizing this essential compound (e.g.
panthothenic acid of coenzyme A).






Genes can be expressed with different efficiencies
Complementary base pairs in the DNA
double helix
Uracil form base pairs with adenine
Differences
between DNA
and RNA
1. The nucleotide in
RNA are
ribonucleotides,
containing sugar ribose
(ribonucleic acid)
rather than
deoxyribose

2. Like DNA, RNA
contains the bases
adenine (A), guanine
(G), and cytosine (C), it
contains the base uracil
(U) instead of thymine
(T) in DNA.
DNA always occur in cells as a double-stranded
helix, RNA is single-stranded.

RNA chain can fold up into a particular shape, just
as a polypeptide chain folds up to form the final
shape of a protein.

The ability of RNA to fold into complex three-
dimensional shapes allows some RNA molecules to
have precise structural and catalytic functions.
Differences between DNA and RNA
RNA can fold into specific structure
From DNA to RNA
The RNA in a cell is made by DNA transcription.

Transcription begins with the opening and
unwinding of a small portion of the DNA double
helix.

One of the two strands of the DNA double helix
then acts as a template for the synthesis of an RNA
molecule.
From DNA to RNA
The nucleotide sequence of the RNA chain is
determined by the complementary base-pairing
between incoming nucleotides and the DNA
template.

The RNA chain produced the transcript is
therefore elongated one nucleotide at a time. It has
a nucleotide sequence that is exactly
complementary to the strand used as the template.
From DNA to RNA
Unlike a newly formed DNA, the RNA strand does
not remain hydrogen-bonded to the DNA template
strand. Instead, just behind the region where
ribonucleotides are being added.

The RNA chain is displaced and the DNA helix
reforms.

Because RNA are copied from only a limited region
of the DNA, RNA molecules are much shorter than
DNA molecule.
Differences in DNA and RNA Replication
Transcription of two genes as observed under
the electrone microscope
Question
How DNA molecule is copied
into RNA (transcription) in
procaryote and eucaryote ?
Transcription Process in Procaryote
Require RNA polymerase holoenzyme

Play a role in transcription process :
- Binding of the polymerase to the DNA
template.
- Initiation of transcription.
- Elongation.
- Termination.

Steps in transcription cycle of bacterial RNA polymerase
1. The RNA polymerase holoenzyme (polymerase core
enzyme plus factor) assembles and locates a promoter.

2. Promoter unwinds the DNA at the position in which
transcription is to begin.

3. The initial RNA synthesis (aborted initiation) is relatively
inefficient.

4. Once RNA polymerase has managed to synthesize about
10 nucleotides of RNA, it breaks its interaction with the
promoter DNA.

5. During the elongation mode, transcription is
highly processive, with the polymerase leaving the
DNA template and releasing the newly transcribed
RNA only when it encounters a termination signal.

Step 6 & 7. Termination signal are typically
encoded in DNA. Many function by forming an
RNA structure that destabilizes the polymerases
hold on the RNA.
Steps in transcription cycle of bacterial RNA polymerase
The transcription cycle of bacterial RNA polymerase
(Procaryote)
The importance of RNA polymerase orientation
Transcription Process in Eukaryotes
Require for a large variety of accessory
proteins, or transcription factors.

Play a role in transcription process :
- Binding of the polymerase to the DNA
template.
- Initiation of transcription.
- Elongation.
- Termination.

Transcription Initiation in eucaryotic cell requires many proteins
Initiation of transription of a
eucaryotic gene by RNA
polymerase
The promoter contains a DNA
sequence called the TATA box, located
25 nucleotides away from the sites at
which transcription is initiated.

Through its subunit TBP, TFIID
recognizes and binds the TATA box.

The RNA polymerase itself
assemble at the promoter.

TFII H uses ATP to pry apart the
DNA double helix.

The site of phosphorylation is a
long C terminal polypeptide tail,
called C terminal domain (CTD).
Summary of the steps leading from gene to protein in
eucaryote and bacteria (procaryote)
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Control of Gene Expression