Regulation of gene

expression

Dr.S.Sethupathy,
RMMC,
Annamalia University
Gene expression

 Gene expression is the process by

which information from a gene is used
in the synthesis of a functional gene
product.
 Products are often proteins
 Non-protein coding genes such as
ribosomal RNA (rRNA), transfer RNA
(tRNA) or small nuclear RNA (snRNA)
genes, the product is a functional RNA.
 Eukaryotes ,prokaryotes
 A gene product is the
biochemical material,
either RNA or
protein, resulting
from expression of a
gene.
Importance of regulation of G.E

By altering gene expression,

organisms can adapt to
environmental challenges.
Transcription control can result in
tissue specific gene expression.
Gene regulation is influenced by
hormones, heavy metals and
chemicals.
Dysregulation of gene regulation
can lead to disease.
Control of gene expression
 Gene amplification
 Gene rearrangement
 Transcription control
 Post transcriptional modification
 RNA stabilization
 Translational control
 Protein modification
 Protein stabilization.
 Positive regulation and negative
regulation.
 A positive regulator, an enhancer or
activator mediates positive
regulation.
 Inhibition of a negative regulator
also results in positive regulation.
 A negative regulator, a silencer or
repressor mediates negative
regulation.
Temporal response
 In type A response, there is increased gene
expression and depends upon the continual
presence of inducing signal. E.g. inducers such as
hormones, nutrients, growth factors.

 In type B response, there is a transient increase in

the gene expression even in the continued
presence of the signal. E.g. it occurs during the
development of an organism.

 In type C response, there is increase in gene

expression that persists indefinitely even after
termination of the signal. e.g. During
differentiation in a tissue or organ, it occurs.
The cistron is the smallest unit
of gene expression.

 A Single mRNA that encodes more than

one translated protein is referred to as a
polycistronic mRNA.

 In prokaryotes, the genes involved in a

metabolic pathway are often present in a
linear array called on OPERON. e.g.
Lac operon.
 An inducible gene- expression increases in
response to an inducer or activator.
 Have low basal rate of transcription.
 Genes with high basal rate are down
regulated by repressors.
 A gene expressed at a constant rate is
constitutive gene
 An inducible gene by mutation -a
constitutive gene- constitutive mutation.
Lac operon
 Operon is a cluster of structural genes
that are expressed as a group and their
associated promoter and operator.
 The lac operon (lactose operon) is an
operon required for the transport and
metabolism of lactose in Escherichia coli
and some other enteric bacteria.
 It has three adjacent structural genes,
lacZ, lacY, and lacA.
 β-galactosidase, lactose permease, and
thiogalactoside transacetylase (or
galactoside O-acetyltransferase),
respectively.
lac Operon Gene Gene Function

I Gene for repressor protein

P Promoter

O Operator

lac Z Gene for ß-galactosidase

lac Y Gene for ß-galactoside permease

Gene for ß-galactoside

lac A
transacetylase
 It is a unit of gene expression.
 It contains a promoter site, operator and
three structural genes.
 Coordinate expression of the three
enzymes and their regulation.
 One large polycistornic mRNA is
transcribed with separate initiation and
stop codons
 Three proteins are produced separately.
 In the promoter site a gene lac I encodes
the repressor Lac I protein.
 It is a constitutive gene.
 It produces four identical subunits of lac
repressor.
 High affinity for operator locus- a region of
double stranded DNA.
 Operator locus is an inverted 21 bp long ,
palindrome.
 5’-AATTGTGAGCGGATAACAATT-3’
 3’-TTAACACTCGCCTATTGTTAA- 5’
The operator locus is present
between promoter and
transcription initiation site (lacZ
gene).
 Lac repressor protein binds to
operator locus
 It prevents the binding of RNA
polymerase to the promoter
site.
So it is a negative regulator.
In the presence of lactose or
isopropyl thiogalactoside
(IPTG), a lactose analog results
in the induction of lac operon
enzymes.
 IPTG induces but is not a
substrate for enzyme and so it
is an example for gratuitous
inducer.
 Small amounts of lactose or
IPTG enter the cell even in the
absence of permease.
Lactose binds to repressor
molecules
This results in the dislodging of
repressor from operator locus.
Now RNA polymerase binds
to promoter and
transcription begins.
Inducer depresses the lac
operon and allows the cell
to synthesize enzymes that
metabolize lactose.
 Bacterium accumulates cAMP when it is
starved for carbon source.
 Lactose is not the preferred carbohydrate
source for E. coli. glucose is utilized first
and the lac operon is turned on. This type
of control is termed catabolite
repression.
 The promoter of the lac operon has two
binding sites. One site is the location
where RNA polymerase binds. The second
location is the binding site for catabolite
activator protein (CAP) and cyclic
AMP (cAMP) complex.
 The binding of the CAP-cAMP complex to
the promoter site is required for
transcription of the lac operon.
 In the presence of glucose or glycerol the
bacteria will lack cAMP to bind to CAP
because glucose inhibits adenylyl cyclase
which converts ATP to cAMP.
 So in the presence of glucose cAMP-CAP is
lacking and RNA polymerase cannot bind.
In the presence of cAMP-CAP
complex which binds DNA upstream
of promoter site and transcription
occurs at maximal levels.
cAMP-CAP is acting as a positive
regulator.
 Maximal expression of lac operon
occurs when glucose level is low
(high cAMP-CAP complex) and
lactose is present.
 If lac I gene is mutated, lac operon
will exhibit constitutive expression.
 Eukaryotic viruses such as helps simplex
virus, HIV can either reside in a dormant
state within the host chromosome or can
replicate and leads to lysis of the host cell.
 Some E.coli harbor a temperate virus
bacteriophage lambda (λ).
 Depending upon the nutritional state, the
lambda DNA either integrate into the host
DNA (lysogenic pathway) and remain
dormant or
 It will replicate until it makes more than
100 copies of virus particles and causes
lysis of the host.
The lytic/lysogenic genetic switching
event is controlled by 80 bp region
in the DNA referred to as the right
operator (OR).
 On the left of OR, there is a gene
for lambda repressor protein (CI).
On the right, there is a gene
encoding for a regulatory protein
Cro.
 When the repressor gene CI is on,
the Cro gene is off and when the
Cro gene is on, the CI gene is off.
Molecular transcriptional switch
 OR is divided into three discrete evenly
spaced, 17 bp cis-active elements (OR1,
OR2, OR3) that have binding sites for the
two regulatory proteins of lambda phage.
 The DNA region between Cro and
repressor genes, have two promoter
sequences.
 One promoter directs RNA Pol to
transcribe rightward direction ie Cro and
other distal genes
 The other promoter directs the
transcription of CI repressor gene in the
leftward direction.
 CI repressor protein forms a dimer and
binds to operator DNA.
 Cro protein is also a dimer that binds to
operator DNA.
 In a lysogenic bacterium, CI repressor
protein preferably binds to OR1 and also
OR2.
 The affinity for OR3 is least.
 The occupation of OR1 by CI repressor
blocks the binding of RNA polymerase to
the rightward promoter and so prevents
the expression of Cro gene.
The binding of CI to OR2 enhances
the binding of RNA polymerase to
the leftward promoter that overlaps
OR3 and enhances transcription of
CI repressor gene.
 So the CI repressor acts as a
positive regulator of CI gene and
negative regulator of Cro gene.
This results in stable state of
dormant lambda bacteriophage ie.
Lysogenic pathway..
 When there is excess CI repressor
protein, it binds to OR3 and blocks its own
transcription till CI level comes down.
 Then repressor CI dislodges from OR3
and transcription continues.
 When a DNA damaging signal such as
Ultraviolet rays, fragments of single
stranded DNA are generated that activate
a specific co-protease coded by a
bacterial gene rec A.
 This rec A protease hydrolyses repressor
proteins and dissociates them from OR1
and OR2.
Now RNA polymerase has access to
the right ward promoter and
transcribes cro gene.
 The cro protein tightly binds to OR3
and turns off the CI gene
transcription.
The Cro gene is expressed now and
repressor gene is turned off.
This is irreversible when Cro gene
protein concentration is high, it
occupies the OR1 and reduces its
transcription..
This is the final stage of lytic cycle.
Both CI repressor protein and Cro
protein bind to DNA using helix-
turn- helix DNA binding domain
(DBD) motifs.
 In proteins, the helix-turn-helix
(HTH) is a major structural motif
capable of binding DNA.
 It is composed of two α helices
joined by a short strand of amino
acids and is found in many proteins
that regulate gene expression.
Regulation of eukaryotic gene
expression
 The chromatin template contributes to
eukaryotic gene transcription control.
 The development of specialized cells
tissues and organs are due to differential
expression of genes.
 For example: DNA containing β- globin
gene is in the active chromatin in
reticulocytes but it is in inactive chromatin
in muscle cells.
Histone covalent modification

 The histone code is essential for gene activity.

 The modifications are dynamic and reversible.
 Acetylation of lysine residues in the amino
terminal tails of histone increases
transcription.
 Acetylation reduces the positive charges in
the tails and reduces the affinity of histone
towards negatively charged DNA.
 This creates additional binding sites for
proteins such as chromatin remodeling
complexes.
 Histone deacetylation catalyzed by
co-repressors will decrease
transcription.
 The proteins that catalyze histone
modifications are called Code
writers.
 The proteins that recognize, bind
and interpret the code are referred
to as Code readers.
 The enzymes that remove histone
modifications are termed Code
erasers.
 Methylation of deoxycytidine (5’me-C)
(sequence 5’-meCpG3’) in DNA of
mouse liver ribosomal genes and some
animal viruses blocks the transcription.
 Many eukaryotic genes have multiple
protein binding DNA elements.
 The binding of transcription factors to
these elements disrupts the
nucleosomes and influences the
transcription.
 Promoter in eukaryotes is more complex.
 It is influenced by many trans-acting
factors which come from other
chromosomes. (So act in trans).
 Covalent modification of DNA, histone and
non-histone proteins stably alters gene
expression patterns without altering the
underlying DNA gene sequence.
 Epigenetics is the study of heritable
changes in chromatin (e.g., DNA
methylation) without involving the change
in DNA sequences. ( it means above
genetics) .
 Two forms of epigenetic signals
ie: cis and trans epigenetic signals.
 A trans signaling event is composed of an abundant
diffusible trans-activators.
 DNA binding trans- activator is transcribed from its
cognate gene (cognate gene- Gene- related or
analogous in nature, character or function).
 This is freely diffusible between nucleus and cytoplasm.
 Excess trans-activator reenters the nucleus following cell
division, binds to its own gene and activates
transcription in both daughter cells.
 Thus it results stable expression of the trans-activator in
both cells.
 trans-acting (trans-regulatory, trans-regulation), means
"acting from a different molecule" (i.e., intermolecular).
 Cis-acting (cis-regulatory, cis-regulation) means "acting
from the same molecule" (i.e., intramolecular).
 A trans-acting element is usually a DNA sequence that
contains a gene.
 This gene codes for a protein (or microRNA or other
diffusible molecule) that will be used in the regulation of
another target gene.
 The trans-acting gene may be on the same
chromosome as the target gene, but the activity is via
the intermediary protein or RNA that it encodes.
 Cis-acting elements do not code for protein or RNA.
 Both the trans-acting gene, and the protein/RNA that it
encodes are said to "act in trans" on the target gene.
 A cis-regulatory element or cis-element is
a region of DNA or RNA that regulates the
expression of genes located on that same
molecule of DNA .(often a chromosome).
 A cis-element may be located upstream
of the coding sequence of the gene it
controls (in the promoter region or even
further upstream), in an intron, or
downstream of the gene's coding
sequence, in either the translated or the
untranscribed region.
 Transmission of epigenetic states.
 Transmission of DNA methylation
patterns after DNA replication.
 Hypothetical model for the maintenance
of a histone-associated epigenetic
signal,.
Maintenance of a chromatin domain via a
secondary signal.
It generates sRNA species that recruit
chromatin-modifying complexes to re-
establish heterochromatic signatures at
the target loci.
o RNA-directed deposition of epigenetic signals.
o Sequence-specific recognition by both sRNAs
and lncRNAs.
o Interaction of an sRNA with complementary sequences on
a nascent transcript.
o Interaction of an sRNA with ssDNA to form a
heteroduplex.
o Recognition of sequence motifs by sRNA in a closed
dsDNA duplex.
o A folded lncRNA recognizes a DNA sequence via complex
surface-mediated interactions.
o A lncRNA acting locally in a co-transcriptional fashion by
being anchored to chromatin by RNAPII.
o RBP, RNA binding protein; adaptor protein;
o CMC, chromatin-modifying complex.
Epigenetics

 Example-the process of cellular differentiation.

 By activating some genes while inhibiting the expression
of others.
 The methylation of mRNA plays a critical role in human
energy homeostasis.
 The obesity-associated FTO gene is shown to be able to
demethylate N6-methyladenosine in RNA.
 subfield of RNA epigenetics.
 1. The modified histones may be carried into
each new copy of the DNA.
 These histones may act as templates.
 These modified histones would ensure that a
differentiated cell would stay differentiated
 Other modifications include acetylation, methylation,
ubiquitylation, phosphorylation and sumoylation.
 2. The second way is the addition of methyl
groups to the DNA, mostly at CpG sites, to convert
cytosine to 5-methylcytosine.
 Highly methylated areas tend to be less transcriptionally
active.
 They also persist from the germ line of one of the
parents into the zygote, marking the chromosome as
being inherited from this parent (genetic imprinting).
Micro RNA and sRNA
 A microRNA (miRNA)-a small non-coding RNA molecule (
about 22 nucleotides) found in plants, animals, and some
viruses.
 Involved in transcriptional and post-transcriptional
regulation of gene expression.
 It has been implicated in heart disease, cancer, muscle
wasting, viral infection, and diabetes. They target mRNA, in
3’-untranslated region(UTR) .
 sRNAs - small (50-250 nucleotides) non-coding RNA
fragments found in bacteria.
 They control gene expression including virulence genes in
pathogens .
 Bind to mRNA and protein targets in prokaryotes.
Prion

 A prion (PrPSc) is an infectious agent composed of

protein in a misfolded form.(Usually DNA, RNA, or both).
 Prion from the words protein and infectious.
 Prions are responsible for the transmissible spongiform
encephalopathies in a variety of mammals
 Bovine spongiform encephalopathy (BSE, also known as
"mad cow disease") in cattle.
 In humans, prions cause Creutzfeldt-Jakob Disease
(CJD), variant Creutzfeldt-Jakob Disease (vCJD),
Gerstmann–Sträussler–Scheinker syndrome, Fatal
Familial Insomnia and kuru.
Genomic imprinting
The father and mother contribute different
epigenetic patterns for specific genomic loci
in their germ cells.
Angelman syndrome and Prader-Willi
syndrome—both can be produced by the
same genetic mutation, chromosome 15q
partial deletion,
the mutation is inherited from the child's
mother or from their father.
This is due to the presence of genomic
imprinting in the region.
Clinical applications

 DNA methylation and cancer

 Variant Histone H2A and cancer
 DNA repair epigenetics and cancer
 Drug development on HAT and HDAC (Histone deacetylase) for
cancer treatment
 Epigenetics can play a role, for example, in the case of Angelman
syndrome and Prader-Willi syndrome.
 Development
 Evolution- Epigenetics can impact evolution when epigenetic
changes are heritable.
 DNA adenine methylation is important in bacteria virulence in
organisms such as Escherichia coli, Salmonella, Vibrio, Yersinia,
Haemophilus, and Brucella.
 In Alphaproteobacteria, methylation of adenine regulates the cell
cycle and couples gene transcription to DNA replication.
 In case of cis epigenetic signal, a gene located on a
particular chromosome carries a cis epigenetic signal
within the regulatory region upstream of the gene
transcription unit.
 This epigenetic signal is associated with active gene
transcription and gene product formation.
 During DNA replication, the chromatin with epigenetic
mark acts as a template and introduces the epigenetic
mark on the newly synthesized unmarked chromatin.
 So both daughter cells contain the epigenetic mark and
ensure the identical expression in both daughter cells.
 Both states can be reversed if either cis or trans
epigenetic signals are removed.
 Epigenetic signals are critical in gene regulation
since any mutations / over expressions contribute
to human diseases.
 Certain DNA elements also enhance or repress
transcription of eukaryotic genes.
 Enhancers enhance initiation at the promoter site.
 Enhancer elements contain multiple binding sites
for trans-activator proteins.
 They may be far off from promoter and they can
act from either upstream or downstream of
promoter region.
 They can act on more than one promoter.
 Examples
 Virally infected cell exerts an antiviral response and
increases interferon production.
 Interferon interacts with neighboring cells and helps in
the inhibition of viral replication.
 Interferon gene is induced by an enhancer element
which is located between -110 and -45 to +1 site.
 This enhancer is composed of form cis elements.
 1. One cis element is bound by NF-kB
 2. Interferon regulatory factor( IRF) a family of trans-
factors bound to a cis element.
 3. A leucine zipper factor
 4. A transcription factor HMGI(Y)
These four factors form the
enhanceosome which increases the
transcription of beta interferon
gene.
There are cis elements that
decrease or repress the expression
of specific genes.
Tissue specific expression may
result from the action of enhancers
or repressors or a combination of
both cis regulatory elements.
 Reporter genes are used to define
enhancers and other regulatory elements.
 Here reporter genes are ligated to regions
of DNA suspected of harboring regulatory
sequences and is introduced in host cell.
 The basal expression of reporter gene will
be in increased if the DNA contains an
enhancer
 Addition of a hormone or heavy metal to
the culture medium will increase the
expression of reporter gene if the DNA
contains a hormone or metal response
element.
 Transfected cells in culture are useful to
identify hundreds of enhancers,
repressors, tissue specific elements,
hormone, metal response elements.
 Combinations of DNA elements and
associated proteins provide diversity in
gene expression.
 Chromatin arrangement in functional units
control pattern of gene expression.
 Some regions are controlled by DNA
elements called to locus control regions
(LCR).
An LCR with associated proteins
controls the expression of a cluster
of genes e.g. globin gene family.

Insulators are DNA elements with

associated proteins that prevent an
enhancer from acting on the
promoter.
Insulators serve as transcriptional
bounding elements.
Locus control regions (LCR)
 They enhance the expression of linked genes to
physiological levels in a tissue-specific and copy number-
dependent manner at ectopic chromatin sites.
 Developmental and cell lineage-specific regulation of
gene expression relies not only on gene-proximal
elements such as promoters, enhancers, and silencers,
but also on long-range interactions of various cis-
regulatory elements and dynamic chromatin alterations.
 Looping model
 LCR section of the DNA loops around to site of the
transcriptional machinery (proteins) and affect the
transcription of that specific gene.
MOTIFS
 In genetics, a sequence motif is a
nucleotide or amino-acid sequence pattern
that has biological significance.
 In proteins, a structural motif is a motif
formed by the three dimensional
arrangement of amino acids which has
biological importance.
 Motifs compose the DNA binding domains
of transcription factor proteins.
 Three unique motifs are helix turn helix,
the Zinc finger, and the leucine zipper.
Proteins that contain motifs
have high affinity to specific
site and low affinity to other
sites of DNA.
Small regions of the protein
make direct contact with DNA
and the rest of the protein
provide surfaces for interaction
of co-activators or co-
repressors.
Helix turn helix
 Bacteriophage lambda Cro
transcription regulator.
Its monomer consists of three
anti parallel sheets and three
alpha helices.
The alpha helices form the DNA
recognition surface.
Zinc finger motif
 TF III A, a positive regulator of 5S RNA gene
transcription requires Zinc for activity.
 It contains nine Zinc ions in a repeating
coordination complex closely spaced to the
cysteine-cysteine residues followed by 12-13
amino acids and later by a histidine- histidine
pair.
 Zinc is coordinated by two cysteine and two
histidine residues.
 The protein containing zinc fingers lie on the
face of DNA helix.
Clinical applications

 A single amino acid mutation in either of

the two Zinc fingers of 1,23(OH2) – D3
(calcitriol) receptor protein results in the
resistance to hormone and causes vitamin
D resistant rickets.
 An ongoing clinical trial is evaluating Zinc
finger nucleases that disrupt the CCR5
gene in CD4+ human T-cells as a potential
treatment for HIV/AIDS.
The leucine zipper motif
 This region of protein forms an α-helix in which there is
a periodic repeat of leucine residues at every seventh
position.
 This occurs for eight helical turns and four leucine
repeats.
 This helps in the formation of separate DNA binding
domain ( DBD) with their target.
 Proteins that regulate transcription have several
domains.
 The DNA binding domain and transactivation domains
are separate and non- interactive.
Clinical applications
 The bZIP (basic zipper proteins) -
transcription factors-its basic region with
the major groove of DNA -hydrogen
bonding, and a hydrophobic leucine
zipper- for dimerization.
 Leucine zipper regulatory proteins - c-fos
and c-jun (the AP1 transcription factor)
 If they are overproduced or mutated in a
vital area, they may generate cancer.
Eukaryotic gene regulation

It occurs at the level of

transcription, nuclear RNA
processing, mRNA stability and
translation.
 In addition, gene amplification
and rearrangement influence
gene expression.
Gene amplification
 During early development, there is an
abrupt increase in the need for specific
molecules such as ribosomal RNA and
mRNA.
 So the number of genes of these
molecules increase by making multiple
copies of ribosomal RNA genes.
 These are generated by a process of
repeated initiations during DNA synthesis
and the amplified genes provide multiple
sites for gene transcription.
Clinical applications
 Methotrexate is an anticancer drug
which inhibits dihydrofolate reductase.
 During long term therapy, cancer cells
develop resistance to this drug by
increasing the number of genes for
dihydrofolate reductase by gene
amplification.
 Oncogenes may also get amplified in
many cancer cells.
Gene rearrangement or somatic
recombination of DNA
 Immunoglobulin IgG has a heavy and a light chain
encoded by several different segments of DNA.
 For example: IgG light chain is composed of
variable (VL), joining (JL) and constant (CL)
domains for a particular subset of IgG.
 There are roughly 76 tandemly repeated VL gene
coding segments
 5 JL coding sequences and 1 CL coding gene for IgG
kappa chain.
 All are located in the same region of the same
chromosome.
Recombination
 So an immune cell has multiple VL, JL, CL segments
to choose from and it helps to develop both
immunologic flexibility and specificity.
 Single VL, JL, CL segments are recombined to
generate a single transcription unit.
 The deletion of unused, remaining segments is
achieved by selective recombination.
 The newly recombined pre- mRNA after removal of
introns by splicing, forms mature mRNA, a single
transcription unit and forms a light chain with VL,
JL, CL segments of one each.
Gene switching
 It is a situation in which a cell or organism stops
expressing one gene or gene group and switches to
expressing a different gene or group of genes.
 e.g: In embryo, hemoglobin is made up of 2 zeta
and 2 epsilon chains.
 By the 6 month of intrauterine life, this is changed to
HbF with 2 alpha and 2 gamma chains.
 After birth, HbF is replaced by adult HbA1 with 2
alpha and 2 beta chains.
 In immune function, during the primary response,
IgM antibodies are produced
 During the secondary response, IgG antibodies are
produced. This is called gene switching.
Alternative RNA processing
 Alternative transcription start sites results
in a different 5’ exon on mRNAs encoding
mouse amylase and myosin light chain.
 Alternative polyadenylation sites in the
IgM heavy chain primary transcript result
in mRNAs that are either 2700 bases long
(µm) or 2400 bases long (µs).
 This results in a different carboxy terminal
and µm protein remains attached to the
membrane of B- lymphotycte whereas µs
protein is secreted.
Alternative splicing and
processing

By alternative splicing seven

unqiue α-tropomyosin mRNAs
are produced in seven different
tissues. How this occurs is not
clear.
 Eg. Deletion of 5’UTR results in 3 to 5 fold
prolongation of half-life of c-myc mRNA.
 The absence of poly (a) tail may cause
rapid degradation of mRNA.
 Histone mRNAs lack poly A tail but they
have a stem loop structure which provides
resistance to exonculeolytic attack.
 Much of this mRNA metabolism may occur
in P Bodies.
Antiviral agents
 Viruses are intracellular and they utilize
the host cell machinery for its replication.
So antiviral agents are targeted at
different steps of virus cycle.
 1. Adsorption and penetration of virus into
host cell is inhibited by antibodies either
passive or active. Neuroaminidase
inhibitors such as Oseltamivir and
Zanamavir are used in the treatment of
influenza.
 Mutations in the virus can lead to
resistance to the drug.
 2. Uncoating of viral nucleic acid is
blocked by amanitidine and is used
against influenza virus.
 3. Synthesis of DNA is inhibited by purine
or pyrimidine analogues.
 Acyclovir is an analogue of guanosine
which gets phosphorylated in the host cell
by the virus specific thymidine kinase.
Then triphosphate form is produced by the
cellular enzymes. This inhibits viral DNA
polymerase.
 Iododeoxy uridine is the first antiviral
drug marketed.
 Ribavirin is an analogue of guanosine. It is
a nucleoside inhibitor. It stops viral RNA
synthesis and inhibits capping of viral
mRNA.
 It is useful against respiratory synctial
virus and viral hemorrhagic fever.
4. Inhibitors of reverse transcriptase
abolish synthesis of RNA. Zidovidine
is a deoxy thymidine analogue.it is
useful against HIV infection.
Didanosine (Di-deoxy inosine) is
also useful in HIV.
5. Protease inhibitors such as
Ritonavir, Saquinavir, Indinavir are
used against HIV infection
Viral infection in patients can be
demonstrated by specific antibodies.
Transduction
 Sometimes virus carries a portion of host
DNA. Subsequent infection of a new host
by this virus may introduce new genes to
the new host.
 This is called viral transduction. Virus acts
as jumping genes.
 During evolution, because of virus the
genetic information from a few organisms
are widely transferred horizontally within a
short period. Thus evolutionary process
has been accelerated by viruses.
Aptamers
 Aptamers (from the Latin aptus - fit, and Greek meros -
part) are oligonucleic acid or peptide molecules that bind to
a specific target molecule. and can be classified as:

 DNA or RNA or XNA aptamers. They consist of (usually

short) strands of oligonucleotides.
 Peptide aptamers. They consist of a short variable peptide
domain, attached at both ends to a protein scaffold.
 Natural aptamers also exist in riboswitches.
 Aptamers can be used for both basic research and clinical
purposes as macromolecular drugs.
 Aptamers can be combined with ribozymes to self-cleave in
the presence of their target molecule.
Structure of an RNA aptamer
specific for biotin.
Artificial nucleic acids

 Nucleic acid analogues are also called Xeno Nucleic Acid.

 Artificial nucleic acids include peptide nucleic acid (PNA)
 Morpholino and locked nucleic acid (LNA)
 Glycol nucleic acid (GNA)
 Threose nucleic acid (TNA).
 Each of these is distinguished from naturally occurring
DNA or RNA by changes to the backbone of the
molecule.
Thank you