At the end of the lecture,

students should be able to:

 Define mobile genetic elements
 Describe the functions of mobile
genetic elements
 Explain the two mechanisms
related to jumping genes
 Differentiate autonomous and non
autonomous transposons
 Explain how transposon shaped
evolution – P elements
 What is transposon?
Organism’s DNA do not remained unchanged.
Can be damaged or mutated / undergo
recombination.
Transposon is a small piece of DNA that can
inserts itself into another place in the genome.
Also known as the jumping genes.
Transposon discovery
Barbara McClintock (1902-
1992; Nobel - 1983)
in1940s planted self
pollinating maize plants
( use the pollen from the
individual to pollinate
the same plant’s flower).
Observed different
characteristics - leaf
pattern, kernel color)
Transposon discovery
How do genes get
dispersed through
genome?
Barbara
McClintock (1902-
1992; Nobel - 1983)
in 1940s initially
ignored this
phenomenon
Transposon discovery
 Found certain mutations in patterns &
markings in leaf & kernel coloration of maize
(corn)
 Some unstable, appearing & disappearing
from one generation to next or even in same
plant
 Concluded some genes had moved from 1 site
to another in chromosome affecting gene
expression
 Called this genetic rearrangement transposition
& the moving genes transposable elements
Transposon discovery

 McClintock worked with what is known as the Ac/Ds system

in maize.
 Through these experiments, McClintock recognized that
breakage occurred at specific sites on maize chromosomes.
 The first transposable element she discovered was a site
of chromosome breakage, aptly named "dissociation"
(Ds).
 Movements of Ds are regulated by an autonomous
element called "activator" (Ac), which can also promote
its own transposition.
Bacterial Transposons
A transposable element moves
from one DNA address to another
Originally discovered in maize,
transposons have been found in
all kinds of organisms
Bacteria
Plants
Humans
Discovery of Bacterial Transposons

Shapiro et al studied phage mutations in 1960s

- phage coat is made of protein and always has
the same volume
DNA is much denser than protein
More DNA in phage, denser phage
gal- phages are denser, suggesting addition
of extra DNAs that inactivate the gal gene.
These extra genes are called insertion
sequences (ISs)
Discovery of transposon

In late 1960s, transposons were found in

bacteria
 They were found to encode a protein
(transposase) that facilitates insertion of
mobile element into the target DNA site;
transposase catalyzes breakage & reunion
of DNA required for insertion
 Transposase catalyzes transposon
excision from donor DNA site &
insertion at target DNA site
 Transposable Element-
sequence of DNA that move from one location in the genome to another,
“ jumping genes”

Called as Insertion Sequence (IS) elements. Contain:

Insertion sequence contained special sequences at the transposon’s
both end (two short inverted repeat flanking seq)
A coding region called transposase:
transposase Set of genes that catalyzes
transposition called the transposase gene
Discovery of Bacterial
Transposons - IS
Cohen discovered that
DNA sequences
flanking the
transposon were
inverted.
One strand can base-
pair and produced a
loop structure.
 Transposition Mechanisms
 Terminal repeats recognized by transposases
& required for transposition into target DNA
(found in both bacteria & eukaryotes)
 Eukaryotic genomes contain large numbers
of transposable elements - ~40% of DNA in
human cell nucleus is derived from
transposable elements
 Vast majority of these elements cannot move
from place to place; they have either been
crippled by mutation or their movement is
suppressed by the cell
Types of transposable elements
1) Transposons
- May/may not leave a copy behind
-Encode gene for transposase

2) Retrotransposons
- always leave a copy behind
-Use reverse transcriptase and an RNA intermediate
- May be evolutionary related to retroviruses (ex HIV)
 Transposition Mechanisms
Transposition frequently involves DNA
replication (CLASS I TE)
1 copy remains at original site
New copy inserts at the new site
Replicative transposition
“Copy and paste”
When it does, non-replicative transposition
(CLASS II TE)
“Cut and paste”
Both strands of original DNA move together from
1 place to another without replicating
 Retrotransposons –
Class 1 TE
Retrotransposons replicate through an
RNA intermediate
Retrotransposons resemble retroviruses
Retroviruses can cause tumors in
vertebrates
Some retroviruses cause diseases such
as AIDS
Before studying retrotransposons, look
at replication of the retroviruses
Retroviruses

Class of virus is named for its ability

to make a DNA copy of its RNA
genome
This reaction is the reverse of the
transcription reaction – reverse
transcription
Virus particles contain an enzyme
that catalyzes reverse transcription
reaction, called reverse
transcriptase
 Retrovirus Replication
Viral genome is RNA,
RNA with
long terminal repeats at
each end
Reverse transcriptase
makes linear, ds-DNA copy
of RNA
ds-DNA copy integrates
back into host DNA =
provirus
Host RNA polymerase II
transcribes the provirus
into genomic RNA
Viral RNA packaged into a
virus particle
1. Copy-and-paste
(Class I TE)
 Cut and Paste – Class II TE
 Studies on bacterial transposition indicate
that this mechanism is mediated by 2
separate transposase subunits that bind to
specific sequences at 2 ends of transposon
 The 2 subunits then come together to form an
active dimer that catalyzes a series of
reactions leading to transposon excision
 Transposase-transposon complex then binds
to target DNA where transposase catalyzes
reactions required to integrate transposon
into its new residence
Mechanisms of transposition
 The repeated sequences flanking a
transposon did not exist before
insertion.
 Transposase cuts the DNA in a
staggered fashion
 Transposon insertion left gaps
 After insertion, gaps are filled.
2. Cut-and-paste
(Class II TE)
Cut-and-paste
(Class II TE)
Mechanisms of transposition
Class II TE: Ex P Elements
 Transposons in human

Thus, many transposons can insert

themselves within center of protein-
coding gene (Alu genes)
Humans - some hemophilias result
from transposon jumping into center
of key blood-clotting gene
~1 in 500 human mutations is result of
transposable element insertion
Some are replicated & DNA copy is
inserted into target site, leaving donor
site unchanged (bacteria)
 Retrotransposons in human and
animals
Several eukaryotic transposons transpose in a
way similar to retroviruses
 TY of yeast
 copia of Drosophila
Start with DNA in the host genome
 Make an RNA copy
 Reverse transcribe it within a virus-like particle
into DNA that can insert into new location
HERVs likely transposed in the same way until
the ability to transpose was lost
 HERV = human endogenous retroviruses
 TY of yeast
Example of a eukaryotic transposable element
is the TY element of yeast.
This element resemble a primitive retrovirus.
A retrovirus is a RNA virus where after being
uncoated in the host cell, converts its RNA to a
DNA copy by the enzyme reverse transcriptase.
The DNA copy of the retrovirus is inserted
into the eukaryotic genome, and it remains
there as a provirus until
a provirus  it is excised and
undergoes transcription to produce new viral
particles.
 TY of yeast
Transposition involves an RNA intermediate
that is generated by transcription of
the TY element
A reverse transcriptase (encoded by
the TyB gene of the element) makes a DNA
copy of the element which is then inserted
into a new site in the yeast genome
Autonomous transposon
Variegation in the color of maize kernels is
caused by multiple reversions of an unstable
mutation in the C locus,
locus responsible for
kernel color
Mutation and its reversion result from Ds
(dissociation) element
Transposes into the C gene
Mutates it
Transposes out again, revert to wild type
Autonomous transposon
Ds and Ac were transposable elements
Ds transposes to C and mutate it,
it causing
the gene to be mutated
Ac was an autonomous transposon, can
induce Ds out of gene C  causing reversion
Ds no not have the transposase gene, so
needed Ac transposase to help it transpose
Transposable Elements in Maize –
reversion of mutation
Role of mobile genetic elements in
evolution
Two schools of thought regarding function of
transposable elements:
1.No function - a genetic parasite; invades &
spreads through organism & offspring, if no
serious adverse effects on ability of host to
survive & reproduce
2.Regardless of origin, once DNA is present in
genome it has potential to be used in evolution
so some think that transposons are a key
mechanism in creating genomic changes that
fuel evolution
 Role of mobile genetic elements in
evolution
 Transposable elements can carry adjacent parts
of host genome with them as they move from
one site to another,
another so 2 unlinked segments of
host genome can be joined to form new,
composite segment
 May be primary mechanism in evolution of
proteins that are composed of domains derived
from different ancestral genes
Complex Transposons
The term “selfish DNA” implies that
insertion sequences and other
transposons replicate at the expense
of their hosts, providing no value in
return
Some transposons do carry genes that
are valuable to their hosts,
antibiotic resistance is among the
most familiar
 Antibiotic Resistance and
Transposons – an example

Donor plasmid has

Kanr, harboring
transposon Tn3 with
Ampr
Target plasmid has Tetr
After transposition,
Tn3 has replicated
and there is a copy in
target plasmid
Target plasmid now
confers both Ampr, Tetr
 P Elements

 Transposable element of Drosophila

melanogaster (P element) is example of
evolution shaped by transposons
 Examination of lab fruit flies and the
natural populations of flies at the start
of the 1900s are devoid of the P element
 In contrast, every member of the
species caught in wild today has
multiple copies of P element
 P Elements

 Thought to have been introduced into single

D. melanogaster within past 80 years,
probably by transmission from individual of
another Drosophila species
 Then it spread rapidly through entire species
population
 Transmission of genetic material from one
species to another, whether between different
fruit flies or different types of vertebrates is
likely mediated by parasites (virus)
 They pick up DNA fragments from one host &
transfer it to subsequent host
 P Elements
The P-M system of hybrid dysgenesis in
Drosophila is caused by conjunction of 2
factors:
Transposable element (P) contributed by the
male
M cytoplasm contributed by the female
allows transposition of the P element
Hybrid offspring of P males and M females
suffer multiple transpositions of P element
causing them to be sterile
 P Elements
P females contained a suppressor of the P
element.
Offspring of either P males or M males with P
females are fertile.
Nowadays P elements is used as mutagenic
element for transpotional studies.