Introduction to

molecular biology
MIC10805
Applications of
Molecular Biology
(2017)
 What is molecular biology?

The central dogma

Put the following words in order...

Protein Translation
Transcription
DNA
RNA
FILL IN THE BLANKS
 What is molecular biology?

The central dogma

‘A unidirectional flow of information’ RNA
replication

Hep C

Transcription Translation
DNA RNA Protein
Reverse
transcription
DNA
replication
RNA
editing
 What is molecular biology?

The central dogma

The conversion of RNA to

DNA via reverse
transcriptase is important
in which technique?
 What is molecular biology?

The central dogma

‘A unidirectional flow of information’ RNA
replication

Transcription Translation
DNA RNA Protein
Reverse
transcription
DNA
replication
RNA
editing
 What is molecular biology?

The central dogma: gene expression

‘The conversion of information encoded by a gene into a protein
product via an RNA intermediary’
Product of gene Product of gene
expression expression
 What is molecular biology?

The central dogma

Where do these processes take place?

1 DNA replication
2
3
Transcription
Translation
5 4
1. Cytoplasm
2. Ribosome
3. Mitochondria
4. Endoplasmic reticulum
5. Nucleus
 The central dogma

Transcription: Gene structure

‘A linear sequence of nucleotides within a DNA molecule which
contains the information necessary to produce a specific
polypeptide* via a messenger RNA (mRNA) intermediate’
 What is DNA?

DNA structure

During your summer job at a biotech company, you isolate a previously unknown virus.
Analysis of its genome reveals that it is composed of a double stranded DNA molecule
containing 14% T (thymine). Based on this information, what would you predict the %C
(cytosine) to be?
14% 28%
36% 72%
 What is DNA?

DNA structure
Why do pyrimidines pair with purines? 23.7 Å
 The central dogma

Eukaryotes vs. Prokaryotes

 The central dogma

Transcription: mRNA processing

In eukaryotic cells, mRNA is processed before leaving the nucleus

A A A AA

5’ Ca
p
 The central dogma

Transcription: Alternative splicing

‘Removal of intronic sequences from pre-mRNA to yield mature
mRNA (exonic regions only) ’
 What is RNA?

RNA structure
Transcription = DNA to RNA

RNA is single stranded. Is uracil a pyrimidine or purine?

In RNA the sugar is ribose
RNA contains the nitrogenous base uracil (instead of thymine).
 The central dogma

Transcription

Sense

Antisense
 The central dogma

Transcription

5’ TTGACGATCCATGAC 3’
Give the sequence of the mRNA transcribed from this
DNA fragment.

Give the sequence of the antisense strand to the DNA

fragment.
 The central dogma

Translation
 The central dogma

Translation

Codon

Translate this mRNA: 5’ AAAGUAGGG 3’

Lys Val Gly
 The central dogma

The central dogma: Translation

5’ AGAGGAAAAGTCCAGGTCTGT 3’
Translate this DNA sequence

But what if AGA is not the first codon……?

5’ AGAGGAAAAGTCCAGGTCTGT 3’
 The central dogma

The central dogma: Translation

What is a reading frame? How many reading frames are there?

5’ AGCGGAGCGGGCGAGCAAGGG 3’

+1 5’ AGCGGAGCGGGCGAGCAAGGG 3’
S G A G E Q G

+2 5’ AGCGGAGCGGGCGAGCAAGGG 3’
A E R A S K

+3 5’ AGCGGAGCGGGCGAGCAAGGG 3’
R S G R A R
 The central dogma

The central dogma: Translation

5’ AGAGGAAAAGTCCAGGTCTGT 3’
Now translate this DNA sequence into the two other reading
frames.
 Proteins

Amino acids
 Proteins

Polypeptides
Using the information about amino acids on the previous slide, highlight the
following areas on the peptide sequence below:
- Hydrophobic region
- Negatively charged region
 Proteins

Polypeptides
Proteins do not remain as chains of amino
acids.
They are folded into higher structures.

Folding requires
chaperones
 Proteins

Polypeptides
Disulphide bridges formed between cysteine's on the same amino acid
chain.
Very strong bonds for maintaining higher structures.
Usually for secreted proteins, why?

Pancreatic ribonuclease A
 Proteins

Protein folding

Newly synthesised proteins fold

into their three-dimensional
structures with the help of
molecular chaperones.

Sometimes proteins are not folded

properly....
 Proteins

Protein folding
Many neurodegenerative disorders involve the accumulation of misfolded
proteins
Mutations

C
Or..
 Mutations

DNA replication
When a cell divides, it makes a copy of its DNA — and sometimes
the copy is not quite perfect......

DNA polymerase
Error rate = 1/100,000 bases.
120,000 mistakes in each cell division.
 Mutations

DNA replication
However, the actual rate of error is 1 in 10 10 bases copied.
Due to the help of an exonuclease domain in the polymerase
complex.

The exonuclease domain recognises and helps correct errors made

by the polymerase.
 Mutations

Mutations can be classified in different ways

Germline mutation Somatic mutation

 Mutations

Mutations can be classified in different ways

Mutations can be classified by their effect on the DNA sequence
or the encoded protein

Effect on DNA sequence:

Substitution/point mutations
Insertion mutations
Can cause
Deletion mutations
frameshift
mutations
 Mutations
Point mutations

CTGGAG ‘Exchange of one nucleotide for another’

CTGGGG Point mutations

Transition Transversion

Purine ↔ Purine Purine ↔ Pyrimidine

Pyrimidine ↔ Pyrimidine

Most common is C  T mutation

There are many known point mutations Thymine
that lead to disease Adenine
If more than 1% of the population carry the
same point mutation in the same position it Cytosine
is known as an SNP Guanine
 Mutations
Small scale mutations
A point mutation may have several consequences (in a protein-
coding region):

CTGGAG
Leu Glu

Silent mutation Missense mutation Nonsense mutation

CTGGAG CTGGAG CTGGAG

CTGGAA CTGGGG CTGTAG
Leu Glu Leu Gly Leu Stop
code for the same code for a different code for a stop
amino acid amino acid codon
 Mutations
Small scale mutations
Insertion and deletion mutations can result in frameshift
mutations

CAG CCA CT ?
 Mutations
Small scale mutations

Name the type of mutation

Name the mutations

below
 Mutations
Large scale mutations

Small-scale mutations: Insertion mutations

CTGGAG
CTGGCAGAG
Insertion mutations – on a larger scale (trinucleotide
expansion) CTGGAG
CTGGCAGCAGCAGCAGCAGCAGAG
Trinucleotide expansion
HTT gene sequence
TTGACGCAGCAGCAGCAGCAG……..CAGATAGAC
L T Q Q Q Q Q Q I D

Polyglutamine tract
> 37 repeats = Huntington’s disease
 Content of the human genome
Tandem repeats: trinucleotide expansion

At present there are 14 trinucleotide repeat disorders that

affect humans.
Nine of these neurologic disorders are caused by an
increased number of CAG repeats.
 Genomes

What is a genome?
Popular consensus is that is it a mix of the words gene and
chromosome
It is all the hereditary material of any particular organism

The genome is encoded by DNA

(except for some viruses which have a genome composed of

RNA)

The genome is all the genetic information, both coding and

non-coding DNA sequences
 The human genome

The human genome consists of two distinct parts....

Nuclear genome Mitochondrial genome

Linear Circular
24 linear chromosomes: 1.657 x 104 nucleotides
22 autosomes and 2 sex
chromosomes. Five to ten
copies per
3.2 x 109 nucleotides
mitochondrion
 The human genome
The mitochondrial genome
Mitochondrial replacement

To create an embryo with

genetic material from three
different people and result in
inheritable genetic
modification.

http://www.bbc.co.uk/news/magazine-28986843
 The human genome
The mitochondrial genome
Mitochondrial (mt)DNA is useful!
mtDNA is not unique to an individual (as it is inherited from the
mother)
So mtDNA can be used to trace maternal lineage
Individuals can be mapped back for generations by analysis of
the mtDNA
In forensics, mtDNA is useful as because each cell has many
copies of it
Samples can be matched with a living relative due to the
maternal inheritance of the whole mitochondrial genome
Jesse James (1847-1882)
Legendary American outlaw, gang
leader, bank and train robber and
murderer
 The human genome
Sequencing of the human genome
The human genome is big…..

Part of a free exhibition.

The first printout of the human

genome to be presented as a
series of books
AGCTTTTCATTCTGACTGCAACGGGCAATATGTCTCTGTGTGGATTAAAAAAAGAGT
 The human genome
DNA packaging
DNA needs to be packaged for protection and to fit into
the cell nucleus.

SEM image: Mag. x7750

DNA associates with

proteins that tightly
wrap it and condense
the structure.
(CHROMATIN)
1st level of DNA packaging is
the nucleosome
 The human genome
DNA packaging

The human karyotype

Karyogram

Chromosomes are
different sizes

G-banding (Giemsa staining)

Autosomes

Staining of condensed chromosomes

Produces a visible karyotype
Chromosomes have characteristic
banding patterns
 The human genome
DNA packaging

Diseases linked to unusual human karyotypes

Aneuploidy
Monosomy
One copy of chromosome (autosome)
Disomy
Two copies of chromosome (autosome)
Trisomy
Three copies of chromosome (autosome)

Cri du chat
Down’s (Down) syndrome
Edwards syndrome
Jacobsen syndrome
Wolf-Hirschhorn syndrome
Angelman syndrome
More information is available on Moodle.
Any questions, please come and see me.