BGDA
Professor Louise Lutze-Mann
School of Biotechnology and
Biomolecular Sciences
Information flow
Replication
Transcription Translation
Replication
Process to replicate both strands of the DNA prior to cell division
– new chromosomes are formed
Raw materials
• DNA to be copied - it will act as the template (pattern) for the
newly synthesized strands
• dNTPs (deoxyribonucleotides – dATP, dCTP, dGTP, dTTP)
• DNA polymerase – the enzyme that will carry out the reaction
• Primer – short piece of RNA required by DNA polymerase to
start the reaction
Parental DNA molecule - The first step is the
each base is paired with separation of the two
its specific partner DNA strands
Each parental strand serves The nucleotides are
as a template for the order of connected to form the sugar-
nucleotides along the new phosphate backbones of the
complementary strand. new strands.
YouTube
Structure of double-stranded DNA
The two DNA strands
are antiparallel - they
run in opposite
directions with
respect to the sugar-
phosphate backbone
catalyses the addition of new nucleotides
2 P released & cleaved to
3 P on incoming nucleotide provide the energy for the reaction
• Must have a 3’OH group to add on to
• Consequently,
• will only elongate DNA in the 5’-> 3’direction
• cannot initiate DNA synthesis unless there is a
primer (short piece of RNA) that contains a 3’OH
for addition of new nucleotides
Leading strand
WHY????
DNA polymerase will only
synthesise DNA in a
5’to 3’ direction
YouTube video: https://www.youtube.com/watch?v=TNKWgcFPHqw
Enzymes involved in DNA Replication
Enzyme (protein) Function
Helicase Unwinds the DNA helix
Single stranded binding protein Keeps the DNA strands separated
Primase Synthesizes the RNA primer
DNA Polymerase III Replicates DNA
DNA Polymerase I Removes RNA primer and replaces with DNA
Ligase Joins the Okazaki fragments
Nucleotide analogues can be used to treat disease
AZT is an analogue
of thymidine:
•blocks DNA
replication
•used to treat AIDS
(HIV) infection
Information flow
Replication
Transcription Translation
Information
storage
Nucleus
Accessing
information
Using
information to
Cytoplasm
make functional
units
Gene expression
§ DNA contains the instructions for how a cell will
function
§ Proteins perform these functions
§ How are the instructions translated from the nucleotide
language of DNA into the amino acid language of
proteins?
§ Via an intermediary - RNA
Overview of Gene Expression
§ DNA is transcribed into
messenger RNA (mRNA),
using base pairing.
§ This mRNA is translated
into protein.
§ The code is a triplet code –
3 nucleotides code for each
amino acid.
Transcription
Process to synthesise a short piece of RNA using a specific
single strand of DNA
Raw materials
• Single strand of DNA to be copied - it will act as the template
• NTPs (ribonucleotides – ATP, CTP, GTP, UTP)
• RNA polymerase – the enzyme that will carry out the
reaction
• Transcription factors – proteins that bind to the DNA to
indicate where the gene sequence commences
DNA vs RNA
Double-stranded Single-stranded
Transcription:
DNA > RNA
Only 1 strand of the DNA is
used as the template for
mRNA synthesis
Transcription occurs
in defined regions of
DNA - genes
Transcription
1. Initiation
•RNA polymerase binds to the
promoter region upstream of
the gene
•DNA strands unwind
•RNA synthesis is initiated by
the RNA polymerase
RNA
polymerase
•The RNA polymerase makes a
complementary RNA copy of
the DNA sequence – it
transcribes the DNA into RNA
•The RNA is called messenger
RNA (mRNA)
Transcription
2. Elongation
•The polymerase moves
downstream, unwinding the
DNA and elongating the RNA
transcript.
•As soon as it is transcribed, the
DNA reforms a double helix.
Transcription
3. Termination
•The RNA transcript is released
•The polymerase transcribes a
terminator sequence which
signals the end of the gene
•The polymerase detaches
from the DNA
Youtube visualisations of transcription:
This is from the Walter and Eliza Hall Institute for Medical Research in
Melbourne
http://www.youtube.com/watch?v=DA2t5N72mgw
This is a very comprehensive overview of transcription
and translation
http://www.youtube.com/watch?v=-ygpqVr7_xs&feature=related
The death cap mushroom
• Amanita phalloides induces death by kidney and liver failure in
more than 50% of those who eat it.
• It produces a-amanitin which binds to RNA polymerase,
preventing transcription and hence inhibiting protein synthesis.
