Al-Farabi Kazakh National University

Higher School of Medicine

Transcription of genetic information and mRNA processing.

Lecturer: PhD Pinskiy Ilya Vladimirovich

LEARNING OUTCOMES
As a result of the lesson you will be able to:

1. Define the terms: transcription, promoter, enhancer, terminator.

2. Describe prokaryotic and eukaryotic RNA-polymerases' structure and functions.
3. Describe phases of transcription, explain the processes happening at each phase and
their importance.
4. Explain the process, importance and difference of Rho-independent and Rho-
dependent termination of transcription.
5. Explain the mechanism of polyadenylation, its importance.
6. Describe the structure of the cap fragment, its synthesis and functions.
7. Describe the splicing mechanism and its meaning.
8. Explain the effect of splicing on gene expression.
Literature
1. Alberts B. et al. Molecular biology of the cell. 6th ed. 2015,p. 179-193.
2. Cooper GM. The Cell: A Molecular Approach. 4th ed., p. 155-175
 • Human chromosomes: 50->250 million base pairs.
• Average gene: 3000 base pairs.
• <5% of DNA codes for protein.
• How to find the genes?

Source: http://commons.wikimedia.org/wiki/Image:DNA_ORF.gif 4
What is a Gene?
• Short stretch of DNA on chromosome.
• Two parts:
Regulatory Coding
region region
DNA
• Information in genes is used to make proteins.
• Two stages: transcription and translation

5
Transcription
• Regulatory region has a binding site for RNA polymerase.
RNA Polymerase

Promoter Terminator

Gene Coding Region

DNA: http://commons.wikimedia.org/wiki/File:Helix84.JPG
RNA Polymerase: http://commons.wikimedia.org/wiki/File:PDB_1sfo_EBI.jpg
copy of coding region
RNA: http://commons.wikimedia.org/wiki/File:Simple_transcription_termination1.svg
messenger RNA (mRNA) 6
 Transcription
• Similar to DNA replication, but different.
• Copies only one of the two strands.
• Makes a copy as RNA, not DNA.

Source: http://commons.wikimedia.org/wiki/File:DNA_transcription.svg
 Gene expression
DNA - RNA - Protein
DNA

Replicatio
DNA Initiation
n
Elongation
Degradatio Transcriptio Processing
n RNA n Export
Translatio Initiation
Degradatio Elongation
n Protein
n Processing
Targeting
 RNA
structure
RNA is single stranded polymer of C, G, A, U

Can have secondary structure but typically not

a double helix
Secondary structures of mRNA
 Types of RNA-
polymerases
DNA acts as a template for RNA synthesis
Synthesis by RNA polymerase -RNA Pol

RNA Pol I makes 28S, 18S, and 5,8S rRNA

RNA Pol II makes mRNA, miRNAs and snRNAs in

nucleus

RNA Pol III makes tRNA and other small RNAs

 Control of
transcription
Transcription initiation by RNA Pol II requires
general transcription factors
 Control of
transcription
Transcription start site usually a TATA box (not always)

TBP (TATA-binding protein) binds, changing DNA

structure.

Recruits transcription factor II proteins (TFIIA, B, …)

then RNA Pol II

Collectively the transcription initiation complex

 Control of
transcription
Since DNA is wrapped around histones, how
does RNA Pol gain access to the promoter?

How does RNA Pol know where to bind?

 Control of
transcription
Transcription initiation also requires:
•activators
•mediators (or co-activators),
•chromatin-remodeling proteins

Activators increase the likelihood of successful transcription initiation

Mediators allow activators to communicate with RNA Pol II

 Transcription factors
• DNA-binding proteins associated with specific
regions on DNA (elements)

• Elements may be as small as 6 nucleotides

• Subtle differences in DNA 3 dimensional

structure alter the ability of proteins to bind
 RNA
processing
Newly synthesized transcripts (mRNA) are processed:

•Splice out intervening sequences (=introns) leaving

expressed sequences (exons)

•“Cap” 5’ end of RNA

•Poly-adenylate 3’ end (Poly A+ tail)

 The CALCA gene Poly “A” sites

5´-- Calcitonin exon CGRP exon --3´

1 2 3 4 5 6
Key features:
• 6 expressed sequences, or exons
• Exon 4 codes for 32 amino acids to form calcitonin
• Exon 5 codes for 37 amino acids to form CGRP

• Two polyadenylation, or poly-A sites, recognized by enzymes that can

facilitate cleavage and poly-A addition at the 3´ end of the corresponding
mRNA

• Intervening sequences, or introns

29
 A closer look at pre-mRNA processing
Let’s consider this gene with three exons, numbered 1-3:

DNA 5´-- --3´

1 2 3

During and after transcription but before splicing, the pre-mRNA

receives two modifications:
RNA 5´-- G AAAAA…..AAAA--3´
--3´
1 2 3

30
 A closer look at RNA splicing
Now let’s splice to remove introns:

5´--G AAAAA…..AAAA--3´
1 2 3

x
The spliceosome recognizes
………………..…… G…………………… sequences at intron/exon
GU……A……AG
AG …
boundaries and cuts introns
out
With both introns removed, our mature mRNA now looks like this:
5´--G AAAAA…..AAAA--3´
1 2 3 31
 Alternative splicing of the CALCA gene
Poly “A” sites
DNA
5´-- Calcitonin exon CGRP exon --3´
1 2 3 4 5 6
Thyroid cells Neuronal cells

RNA
Calcitonin
5´-G exon A..A-3´ 5´-G CGRP exon A..A-3´
1 2 3 4 1 2 3 5 6

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 Translation and post-translational processing
Poly “A” sites
DNA
5´-- Calcitonin exon CGRP exon --3´
1 2 3 4 5 6
Thyroid cells Neuronal cells

RNA
Calcitonin
5´-G exon A..A-3´ 5´-G CGRP exon A..A-3´
1 2 3 4 1 2 3 5 6
(Other peptides corresponding
to other translated exons are
Calcitonin CGRP (37
removed during post-
(32 amino amino acids)
Protein translational processing and not 33
acids)
Thanks for attention!