MOLECULAR

BIOLOGY IN
MEDICINE
A. A. Osuntoki, Ph.D.

Molecular biology involves the study of DNA and
RNA, the role of these molecules in biological
systems and how interaction(s) with other
molecules and / or environmental factors affect
their function.
The primary role of DNA is to carry genetic
information.
There are various forms of RNA and they have
various roles in protein synthesis.
WHAT IS GENETICS ?
The study of heredity and variations.
The study of how characteristics are
passed from parents to offspring.
Also studies how new traits arise.

GENETICS AND CLINICAL PRACTICE
Genetic variation is linked to many human
diseases.
Some 4000 genetic diseases have been identified
Genetics is becoming an integral part of health
care in all medical specialties.
Clinician are at the interface of science and care
Lack of genetic training is a common problem for
health professionals.


Knowledge of genome structure and
function coupled with advances in DNA
technology are aiding the development of
novel diagnostic and treatment strategies.
There is a need to understand the basics
as genome technologies move from the
bench to the clinic.
THE CHROMOSOME
A nucleoprotein complex
Contains the genetic material
Polygenic
Consists of DNA and basic proteins
The protein kind and types may vary in
different cell types
Twenty three (23) pairs in humans


THE GENE
Contained on the chromosome
The unit of heredity
Finite sequence of nucleotides
In eukaryotes, each cell contains the full genetic complement
(exception: mature rbc)
Made up of DNA in most life forms
Contains information for the blueprint for the organism
Sequence of DNA capable of independent expression
The major function is to specify the sequence of amino acids in a
protein
Determines the structure of a protein
Must be expressed to perform its biological roles.
Specific genes determine hereditary traits




DEOXYRIBONUCLEIC ACID (DNA)
Is the store of genetic information in all life forms except
some viruses where RNA is the genetic material
Double stranded helix
The two strands are anti-parallel
The structure is stabilized by hydrogen bonds between
base pairs (AT, GC)
The double stranded chain structure ensures
information retrieval in case of damage to a strand.
In the cell DNA serves as a template for its own
replication and as a template for RNA synthesis.

BASE PAIRING IN DNA

DNA STRUCTURE

THE GENETIC CODE
Genetic information is organized into genes or cistrons
The message is encoded in a triplet of bases called codons
Codons specify the amino acids to be incorporated into a
growing protein chain
Codons also specify chain termination
There are 64 codons
61 codons specify amino acids
3 codons specify chain termination
The code is degenerate
The code is universal


RIBONUCLEIC ACID (RNA)
A close chemical relative of DNA.
The major difference is the absence of oxygen
at the 2
I
position of the sugar-phosphate
backbone of DNA which is present in RNA.
RNA is single stranded while DNA is double
stranded.
Present in several different forms in living cells
* messenger RNA (mRNA)
* transfer RNA (tRNA)
* ribosomal RNA (rRNA)


GENE EXPRESSION
A gene is said to be expressed when the
gene product is being produced.
The flow of genetic information is depicted
below:

DNA RNA PROTEIN
i ii

The implication is that DNA makes RNA which in turn makes
proteins.
This is what is generally referred to as the Central dogma of
gene expression.
The formation of daughter DNA strands is called replication.
This process is semi conservative i.e. a strand in a new pair
is newly formed while the other is inherited.
The transfer of genetic information to RNA (i) is called
transcription. All RNA types are transcribed from DNA.
The conversion of the genetic information into a polypeptide
(ii) is called translation.

REPLICATION
Replication is semiconservative.
Starts at a unique origin and proceeds in
opposite directions.
Both strands serve as templates.
DNA synthesis is primed by RNA.
One strand is synthesized discontinuously.
DNA polymerase takes instruction from the
template.
DNA polymerase corrects mistakes during
replication.

TRANSCRIPTION
Only one strand of DNA is transcribed in a
particular region of the genome.
Direction of synthesis is 5
1
3
1
like that of DNA.
Transcription is initiated at promoter sites on the
DNA template.
RNA polymerase takes instructions from a DNA
template.
Many RNA molecules undergo post-
transcriptional modifications.


TRANSLATION
The conversion of the information in a gene into
a protein molecule.
The process of reading the genetic code.
Takes place in the ribosome.
mRNA is the template.
The tRNAs carry amino acids in the activated
form to the ribosome for incorporation into a
nascent chain.
AUG is the start codon while UUA, UAG and
UGA are stop codons.


PHENOTYPES AND
GENOTYPES
Phenotypes : All the observable
characteristics of an individual
Traits: refer to the single characteristic of the
individual
Genotypes: The genes that make up the
chromosomes showing a characteristic trait
Genes contain the actual code for traits
Alleles: variations of a gene that relate to the
same characteristic are called alleles



GENETIC DISORDERS
Genetic disorders can result from:

Abnormal chromosome numbers
Changes in the reading frame caused by
deletions and insertion
Changes in nucleotide sequence
MUTATION AND DISEASES
The information carrying function of DNA
requires the protection of the integrity of the
molecule.
It is apparent that faithful transmission
(copying) of the information at each
generation is essential for preserving the
characteristics of organisms.
.

Mutation contd
A nucleotide sequence alteration may
change the composition of nucleotides in a
codon, or affect the way the gene is read.
This may result in the incorporation of an
incorrect amino acid into the protein .
A mutation may have significant effects on
the organism or may be harmless

mutation contd..
An altered amino acid sequence may greatly affect the ability
of the protein to perform its biological role.
The chemical and physical features (including the structure)
of a protein result from the arrangement and composition of
amino acids in the chain.
The function of biological molecules is intrinsically linked to
the structure. Thus, the replacement of even a single amino
acid with another may diminish the capacity of a protein to
function or inactivate it.
The end results are genetic defects which may manifest with
negative impacts on the health of individuals.



Alterations in the nucleotide sequence results in
mutation

There are two broad classifications of mutation
- Point mutation (single base substitution)

- Frame shift mutation (indel)

Mutation contd

If one purine [A or G] or pyrimidine [C or T] is replaced by the
other, the substitution is called a transition
If a purine is replaced by a pyrimidine or vice-versa, the
substitution is called a transversion

Samesense: The mutation results in the formation of a codon
that is a synonym of the wild type
Missense: The mutation alters the codon to produce an
altered amino acid in the protein product
Nonsense: The mutation changes a codon for an amino acid
into a stop codon
POINT MUTATION
Insertion
Deletion

FRAMESHIFT MUTATION
GENETICS AND DRUG THERAPY
Virtually every human ailment, except
perhaps trauma, has some genetic basis.
Genetic variation can affect the ability of
individuals to respond effectively to
medication.
This may result in adverse drug reaction or
poor drug response.
Genetic polymorphism is important not only in
determining the predisposition to diseases but
also for determining an individuals ability to
metabolize different therapeutic agents given for
alleviating disease severity.
PHARMACOGENETICS: Examines the link
between genes and drug response.
PHARMACOGENOMICS: Examines the inherited
variations in genes that dictate drug response and
explores ways in which these variations can be
used to predict a patients response to a drug.

GENETICS AND DRUG THERAPY
PHARMACOGENOMICS
The aim is to improve the overall process of
drug development and develop products
that will benefit the largest population.
The ultimate goal is to provide new
strategies for optimizing drug therapy
based on each patients genetic
determinants of drug efficacy and toxicity.
PROSPECTS OF PHARMACOGENETICS
Personalized medications
More powerful drugs
Better, safer drugs the first time
More accurate determination of appropriate
drug doses
Improvements in the drug discovery and
approval process
Decrease in health care costs.
CLONING
Transfer of a gene from one cell to the other.
Method for purifying, propagating and amplifying a
specific DNA sequence or gene.
The cloned fragment is placed under the genetic
expression machinery of a suitable host cell.
The extraneous DNA is faithfully copied.
The cloned DNA is inherited by the clone cell line.
Possible because of the universality of the genetic
code.

DNA AMPLIFICATION

DNA amplification protocols enable the scientist to
increase the copy of number of a gene or DNA of
interest.
Unlike in cloning, the chromosome does not have
to be fragmented and extensive purification may
not be carried out.
The most commonly used amplification protocol is
the polymerase chain reaction (PCR).
PCR is a primer directed in vitro DNA
amplification protocol which requires knowledge of
the nucleotide sequences of the flanking regions
of a gene / region of interest for primer synthesis.
POLYMERASE CHAIN REACTION
(PCR)
PCR is a primer directed enzymatic
amplification process
It enables the amplification of specific DNA
sequences
The target sequence is defined by the primers
The sequences of the flanking regions to the
target site must be known
The primers are complementary to the flanking
regions
PCR: A POWERFUL TOOL
PCR is one of the most powerful laboratory tools
available
It possesses a unique combination of attributes
It can be carried out with comparative ease
It is sequence specific
It is sensitive
It is flexible
It is relatively cheap
Permits rapid amplification of template DNA

APPLICATIONS OF PCR
Due to the simplicity and versatility, PCR
has become ubiquitous and with a wide
range of general applications
The application will be considered under 3
broad headings:
Clinical applications
Forensic applications
Research applications
CLINICAL APPLICATIONS
There are three primary diagnostic applications
Genotyping
Detecting unknown mutations
Detecting pathogens
Other areas of possible use include:
Tissue typing
Blood typing
OTHER USEFUL TECHNIQUES
GENE THERAPY
Clinical intervention involving the transfer of a gene into a
patient with the aim of correcting a genetic disease.

ANTISENSE TECHNOLOGY
This utilizes oligonucleotide sequences designed to be
complementary to a specific gene sequence to alter gene
expression. The aim is to prevent protein expression from a
targeted sequence.


GENOMICS AND PROTEOMICS


Genomics and proteomics seek to decipher the
fundamental biology of living organisms and to use
the information to enhance the quality of life
The study of genomes is called genomics
An organisms complete set of DNA is known as its
genome.
A genome is the total genetic complement of an
organism.
DNA is the molecule that contains the instruction for
the structure and function of nearly every life form.
Genetic information is determined by the sequence
of nucleotides in a DNA molecule.

GENOMICS AND PROTEOMICS CONTD.
Proteins make up the body structures and are involved in numerous
cellular processes.
Alterations in the sequence of nucleotides in a DNA (mutation) can
generate abnormal proteins with altered functions leading to
diseases.
Sequencing of DNA is a useful technique of detecting variations and
mutations which may play a role in the development/ progression of
a disease and in drug response and adverse drug reactions.
The complete DNA sequences of the genomes of many organisms
are available.
The genomic data along with proteomics and the tools of
bioinformatics and computational biology enable detailed studies of
genes and an understanding of the properties of the encoded
proteins.


CONCLUSION
A thorough understanding of the molecular basis of
heredity will ultimately reveal how various elements
interact to affect the human body in health and
disease. There is a promise of a tomorrow where
diagnosis will be exact and the drugs prescribed
more effective and far less likely to induce adverse
drug reactions.