Gene Technology

Lecture 8 – Chapter 7

Mobile DNA Sequences in the Genome

Barbara McClintock (nobel prize

winner 1983)
Found in the late 1940s -> genetic
elements in maize can direct their own
movement within the genome

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Lecture 7 – Chapter 7

Mobile DNA Sequences in the Genome

Mobile Elements:

-> can be found in all organisms

-> 50% of human genome
-> interfere with gene expression
-> generating mutations (-> Evolution)
-> reorganize genomic structure

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Mutation can be caused by insertions:

Mutation in the gal operon was moved

to λ phage

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Mobile elements -> Intergration into genome

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Principle of mobile elements -> Intergration into genome

Transposase-> Enzyme coded on mobile elements

-> resposible for excition and integration

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Insertion Element (IS): -> most simple mobile elements
-> found mainly in bacteria

IS element is cut out and leaves behind flanking repeats (non replicative)
-> orginal target sequence are increased after jumb

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Insertion Element (IS):

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Transposons (TN): -> More complex mobile elements
-> found in all organisms
Transposons:

Transposons carry
frequently antibiotic
resistence genes

Transposons can jump

from genome to phage or
conjugative plasmid -> gene
transfere to other
bacteria (problem with
antibiotica resistent
bacteria)

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Transposons

Composite Transposon:
-> Insertion elements on both
end of the transposon (one
active transposase – one
inactive transposase)

Simple Transposons:
-> Inverted repeats on both
ends
-> Transposase and Resolvase
-> responsible for transfer

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Transposons

Simple Transposon
(Tn3):
-> Replicative
transposition -> after the
jump both have a copy of
the transposon

Composite Transposons
(Tn10):
->non replicative
(conservative) -> like
tranposition of IS
elements

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Transposons

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Transposons

Simple Transposon
(Tn3):
-> Replicative
transposition -> after the
jump both have a copy of
the transposon
Transposase: responsible
for excition and transfer
Resolvase: responsible for
resolution -> replicative
transfer

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Transposons

An R plasmid may contain several transposons carrying

(resistance genes)
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Transposons

Composite transposons:

-> non replicative transfer

Ac element (first discovered in

maize)
-> Activator (avtive transposon)

Ds element (first discovered in

maize)
-> Dissociation
-> passive transposon-> needs Ac to
move

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Transposons

Composite transposons:

-> non replicative transfer

Ac element (first discovered in

maize)
-> Activator (avtive transposon)

Ds element (first discovered in

maize)
-> Dissociation
-> passive transposon-> needs Ac to
move

Deletions in Ac element -> gives Ds

element -> inactive transposase ->
needs Ac to move

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Mechanism of Ac or Ds tranfer

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Transposons
Transposition has influence
on phenotype of maize

First recognized by
Barbara McKintock (first
transposon found)

Movement of Transposon
during development of fruit
-> pattern in pigmentation
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Transposons

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Transposons

Transposons are activated by

hemimethylated DNA -> right
after replication when second
strand is not yet methylated

When non replicative transposon

(Tn10) jumps it leaves a double
strand break behind -> repaired
by using other stand (with
transposon in) as template
(repair pathway) -> in the end
both strand have the
transposon in 2 times

Not only both cells after

replication have the transposon
-> also number of transposons
increased!!!!
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Retrotransposons are similar to retroviruses

RNA intermediate + use

of reverse transcriptase

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Retrotransposons are similar to retroviruses

RNA intermediate + use

of reverse transcriptase

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Transposons versus Retrotransposon

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Retrotransposon
There are 2 groups:

1. LTR (long terminal repeats) retrotransposons (viral like)

Carry strong promoters
If transposon inserted 5’ to silenced
gene -> activates gene expression of
these genes
Retrotransposons can cause cancer!!
Promoter
necessary for
transposition

2. Non-LTR retrotransposons (polyA)

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Retrotransposon

Protein coded for :

Integrase + Reverse
transcriptase

Carry strong promoters for RNA

ploymerase
If transposon inserted 5’ to silenced
gene -> activates gene expression of
these genes
Retrotransposons can cause cancer!!
Retroviruses can also cause cancer!!

ORF1: RNA binding protein

ORF2: Endonuclease + reverse
transcriptase

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Retrotransposon

Demonstration that
retrotransposon transfers
through RNA intermediate

Ty is yeast LTR retrotransposon

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Non-LTR Retrotransposon

ORF1: RNA binding protein

ORF2: Endonuclease + reverse
transcriptase

Promoter
necessary for
transposition

mRNA transcript with poly A

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Non-LTR Retrotransposon

Transposition -> target-primed reverse transcription

Reverse transcription commonly stops before element

has been fully transcribed
-> a lot of truncated L1 elements

Alu elements: 300bp repetitive sequences (11% of human

genome) -> belong to SINEs (do not encode their own
reverse transcriptase -> nonautonomous transposons

SINEs are probably trancated LINEs (share same 3’

sequence)

Alu elements are activated by LINEs

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Types of transposable elements in the human genome

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Human genes contain many transposable elements

Important for survival -> Exon/Intron concept of a gene

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Transposons are mainly responsible for differences in genome size

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Transposons can cause Mutations
Transposable
elements at work in Result of transposition or excision -> not always clean excited -> has an
snapdragon effect on enhancer -> different expression level

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Transposons can cause Mutations
Transposons and retrotransposons can induce mutations by inserting near or within genes.
-> transposon-induced mutations are relatively stable, because the sequence at the
insertion site is retained as they transpose via the replication mechanism.

Most mutations caused by transposons are deleterious -> L1 insertion into hemophilia A

Retrotransposons can be responsible for genetic disease (34 diseases identified) -> hemophilia,
muscular dystrophy, …
Development of genetic disease can happen by transposon jumb in the embryo!!! 34
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Transposons generate diversity for evolution

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Transposons generate diversity for evolution

Can affect genes (delection, insertion, exchange), regulatory elements

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Transposons generate diversity for evolution
Can affect genes (delection, insertion, exchange), regulatory elements

Mutant receptors -> cannot remove cholesterol from blood

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Transposons can be silenced

Organisms with high transposition (Drosphila) can silence transposition

-> P element encodes not only transposase but also an inhibitor to transpostion (Protein alters
slicing of transposase -> inactive transposase)

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Transposons can be silenced

If female has P element ->

oocytes (eggs) with inhibitor
produced -> normal flies

If female has no P element ->

oocytes have burst of
transposition in embryo ->
steril offspring

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