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CHAPTER 7

Mobile Genetic elements

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?
Organisms 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 plants flower).
Observed different
characteristics - leaf
pattern, kernel color)

Transposon discovery
How do genes get

dispersed through
genome?
Barbara
McClintock (19021992; 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 theAc/Dssystem

in maize.
Through these experiments, McClintock recognized that
breakage occurred at specific sites on maize chromosomes.
The first transposable elementshe discovered was a site
of chromosome breakage, aptly named "dissociation" (Ds).
Movements of Dsare regulated by anautonomous
elementcalled "activator" (Ac), which can also promote
its owntransposition.

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 Bacterial Transposons

Insertion sequence contained special

sequences at the transposons both end


Set of genes that catalyzes transposition called
the transposase gene

Discovery of Bacterial
Transposons
Cohen discovered that
DNA sequences
flanking the
transposon were
inverted.
One strand can basepair and produced a
loop structure.

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.

Mechanisms of transposition

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

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

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

Transposition Mechanisms
Transposons are sometimes called jumping genes,

DNA doesnt always leave one place for another


When it does, nonreplicative transposition (CLASS
II TE)
Cut and paste
Both strands of original DNA move together from 1 place

to another without replicating

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

1. Cut-and-paste (Class II TE)

Transposition Mechanisms

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

1. Cut-and-paste (Class II TE)

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

Retrotransposons
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

2. Copy-and-paste (Class I TE)

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, 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

Transposons in human
Thus, many transposons can insert

themselves within center of protein-coding


gene
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
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

theTYelement of yeast.
This element resemble a primitive retrovirus.
Aretrovirusis a RNA virus where after being
uncoated in the host cell, converts its RNA to a
DNA copy by the enzymereverse transcriptase.
The DNA copy of the retrovirus is inserted into
the eukaryotic genome, and it remains there
as aprovirusuntil 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
theTYelement
A
reverse
transcriptase
(encoded
by
theTyBgene 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, 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, 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, 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

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

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.

End of lecture today