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Transcription

Learning Outcomes
At the end of this topic, students should:
• Understand DNA transcription
• Understand RNA translation

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1. Introduction
• Transcription is the transfer of the genetic information
from the archival copy of DNA to the short-lived
messenger RNA (mRNA)

How Cells Decode and Use the

Information in their Genome:
From DNA to Protein
(Eukaryotic Cell)

• RNA (i.e. mRNA) has the same “language” as DNA

Figure 6-2 Molecular Biology of the Cell (© Garland Science 2008)

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Transcription: Securing the valuable data

“DNA”

“RNA transcripts”

• Many identical RNA can be made from the same gene, each RNA
molecule can direct the synthesis of many identical protein
molecule
• Each gene: transcribed-translated with different efficiency – vast
quantities of proteins

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How do DNA carry genetic codes?

• The “language” of DNA only has four
alphabets – A, T, C and G
• These four alphabets (CGAT) can make up
“genetic words” but the words in genetics only
has three alphabets – codons
• These combination of three nucleotide
encodes for a specific amino acids.
• The amino acids produced will form proteins
that will then be used for biological processes.

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DNA transcription
• DNA transcription is the process where by the RNA is
formed from the DNA that carries genetic codes.
– Protein production
– Formation of non-coding RNA (rRNA, tRNA,
miRNA, snRNA)
• The RNA produced by transcription is what we call
the messenger RNA (mRNA)
• Messenger RNA - synthesized from coding region
(exon)
• Once messenger RNA is produced, it will then exit
the nucleus (in eukaryotes) and enter the cytoplasm
for translation process

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Transcription in Prokaryotes

• Transcription is catalyzed by RNA polymerase

which makes RNA using DNA as a template.

– E. coli RNA polymerase – most well-studied

– molecular weight about 500,000 Dalton
– four different types of subunits: α, β , β’, and
σ
– the core enzyme is α2, β, β’
– the holoenzyme is α2, β, β’, σ

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– the role of the σ subunit is recognition of the

promoter locus; the σ subunit is released after
transcription begins
– From these two DNA strands, the one that
serves as the template for RNA synthesis is
called the template strand or antisense strand;
the other is called the coding strand or sense
strand
– the holoenzyme binds and transcribes only the
template strand

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• Transcription of DNA occurs in four main stages:

1. binding of RNA polymerase to DNA at a promoter,
2. initiation of transcription on the template DNA
strand,
3. subsequent elongation of the RNA chain, and
4. eventual termination of transcription, accompanied
by the release of RNA polymerase and the
completed RNA product from the DNA template.

• RNA polymerase moves along the template strand of

the DNA in the 3-prime (3`) to 5-prime (5`) direction,
and the RNA molecule grows in the 5` to 3` direction.

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How does RNA polymerase know

where to begin transcription?

• The DNA promoter region in prokaryotes is a

stretch of about 40 base pairs adjacent to and
including the transcription startpoint.

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• The essential features of the promoter are;

– startpoint (designated +1 and usually an A),
– the six-nucleotide -10 sequence,
– the six-nucleotide -35 sequence.
• The two key sequences are located
approximately 10 nucleotides and 35
nucleotides upstream from the startpoint.
• The number of nucleotides separating the
consensus sequences from each other and
from the startpoint are important for promoter
function, but the identity of these nucleotides
is not.

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• During elongation, RNA polymerase binds to about 30

base pairs of DNA
• Termination of transcription requires a termination
sequence that triggers the end of transcription.
• Two classes exist, rho dependent and rho
independent.
– In rho independent termination, a short complementary
GC-rich sequence (followed by several U residues) will
form a "brake" (haipin loop) that will help release the RNA
polymerase from the template (at the weakly poly-U
stretch).
– In rho dependent termination, binding of rho to the mRNA
releases it from the template.

Rho independent termination

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Overall process

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Regulation - prokaryotes

The control of transcription is largely responsible for
the control of expression of genes; done by

alternatives factors – different s factors for
different genes

Enhancers/silencers upstream of promoters

Transcription factors bind to it

Operons – group of genes that are controlled
together

Inducer will determine if the gene will be
expressed or not (i.e. no inducer – gene not
expressed)

transcription attenuation eg. Trp operon

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Regulation : Lac Operon

Regulation : Trp Operon

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Transcription in Eukaryotes

Transcription in eukaryotes
• Although transcription in eukaryotes is similar to that
in prockaryotes, the process appears to be complex.
• Instead of one RNA polymerase, there are three (RNA
Polymerases I, II, and III) involved in eukaryotic
transcription.
1. RNA polymerase I (localized to the nucleolus) transcribes
the rRNA precursor molecules.
2. RNA polymerase II produces most mRNAs and snRNAs.
3. RNA polymerase III is responsible for the production of
pre-tRNAs, 5S rRNA and other small RNAs.
• The mitochondia and chloroplasts have their own RNA
polymerases.

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Three classes of eukaryotic promoters

1. RNA polymerase I: The promoter for RNA
polymerase I has two components:
• a core promoter (surrounding the startpoint)
• an upstream control element. After the binding
of appropriate transcription factors to both
sites, RNA polymerase I binds to the core
promoter.

2. RNA polymerase II: The typical promoter for RNA

polymerase II has a short initiator sequence,
consisting mostly of pyrimidines and usually a TATA
box about 25 bases upstream from the startpoint.

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3. RNA polymerase III: The promoters for RNA polymerase III

vary in structure but the ones for tRNA genes and 5S rRNA
genes are located entirely downstream of the startpoint,
within the transcribed sequence.
• In tRNA genes, about 30-60 base-pairs of DNA separate
promoter elements;
• In 5S rRNA genes, about 10-30 base-pairs promoter
elements.

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• General transcription factors and the polymerase

undergo a pattern of sequential binding to
initiate transcription of nuclear genes.
– TFIID binds to the TATA box followed by
– the binding of TFIIA and TFIIB.
– The resulting complex is now bound by the
polymerase, to which TFIIF has already attached.
– The initiation complex is completed by the addition of
TFIIE, TFIIJ, and TFIIH.
– After an activation step requiring ATP-dependent
phosphorylation of the RNA polymerase molecule, the
polymerase can initiate transcription at the startpoint.

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• Termination signals end the transcription of

RNA by RNA polymerase I and RNA
polymerase III without the activity of hairpin
structures as seen in prokaryotes.
• mRNA is cleaved 10 to 35 base-pairs
downstream of a AAUAAA sequence (a poly-A
tail signal).

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Post-transcriptional modification

• Messenger RNA in eukaryotes requires

excessive processing
– Capping of 5’ end
• 5 prime "cap" (a guanosine nucleotide
methylated at the 7th position) – to give
stability to mRNA
• Protect from nuclease degradation
– Polyadenylating - Adding poly-A
• Protect from nuclease degradation
– Splicing – removal of introns (non-coding)
from pre-mRNA
mRNA

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Figure 6-21 Molecular Biology of the Cell (© Garland Science 2008)

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3.3 Additional of Poly-A tail

• Poly-A polymerase (PAP) adds 200

A (adenosine) nucleotides at opened
3’OH end of mRNA
• The cap and poly A tail protect the
mRNA from enzyme degradation
and assist its attachment to the
ribosome

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END OF LECTURE

ACKNOWLEDGEMENT:
AZANI SALEH

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