DYNAMIC MUTATIONS

…now the defect found in

Arabidopsis thaliana

Sandep Yadav
 DYNAMIC MUTATION ??
 Unstable heritable element where probability of mutation is
function of no. of copies of mutation.

Unique Properties:
 Expanding, Unstable DNA repeats.
 Repeats are usually polymorphic in copy number, with mitotic and meiotic
instability.
 Copy number related to the severity and/or age of onset
 Imprinting effects
 Reverse mutation - The mutation can revert to normal or to a premutation
carrier state.
 Founder effects are observed in association with dynamic mutation as a higher
frequency of particular allele in the affected population.
TYPES OF DYNAMIC MUTATION
 Genic location of disease-associated repeats

Expanded repeats that cause disease are found in many

different genic locations
How do expanded repeats cause disease?

 Gain of function:
• Repeats in protein coding sequences - toxic proteins
e.g. polyglutamine disease.
• Repeats in RNA coding regions - altered RNA function
e.g. splicing in Myotonic dystrophy.

 Loss of function:
• Repeats in non-coding regions - reduced transcription
or translation
e.g. fragile X syn. & IIL1 in A. thaliana
 Processes involved in repeat expansion

Repeat expansion can take place in the germ line or in the somatic cell
Genetic Variation in A. thaliana’s Bur-0 strain
 Strains were grown at 27 C and short day condition
 All except Bur-0 strain showed normal growth.
 Bur-0 strain showed abnormal growth and no flowering.
 Phenotype was named as “irregularly impaired leaves”(iil).
 At 23 C this defect disappeared but worsen at 30 C.

Pf-0 at 27 C Bur-0 at 27 C Bur-0 at 23 C

 Genetic basis of iil defect

Location of expanded repeats RNA level of At4g13430

 Interval mapping by PCR revealed expansion of GAA repeats in Chrm. 4

 At 23 C RNA level didn’t differ between genotypes while
 RNA level decreased by 50% in Bur-0 strain at 27 C
Overexpression of mature Bur-0 transcript

Bur-0 plant at 27 C (5 week)

IIL expression w.r.t. Overexpression

Bur-0 plant harboring 35S:IIL1-Bur-0 transgene

 GAA repeat and somatic variability
 Phenotypic revertant showed the variability in the repeat no.
 EMS promotes the contraction of triplet repeats.
 After treatment 156 phenotypic revertants, 27 had reduced triplet copy no.

DNA blot analysis of IIL1 triplet repeat PCR analysis of IIL1 repeat.
containing region
 CONCLUSION:
• Bur-0 strain provided a genetically tractable model for TNR studies.

• Phenotypic revertants highlight the potential of IIL1 repeats for further study.

• The molecular mechanisms underlying the repeat disorders involve either loss
of protein function or gain of function at the protein or RNA level.

• Repeat instability involves the formation of unusual DNA structures during DNA
replication, repair and recombination.

• Simple repeats can be associated with phenotypic variability of evolutionary

significance.
Suggested reading:
•Kumar, S., Todesco, M., Schneeberger, K., Harilal, R.,
Balasubramanian, S., Weigel, D. (2009). A Genetic Defect Caused by a
Triplet Repeat Expansion in Arabidopsis thaliana. Sciencexpress 1126: 1-
10.
•Al-Mahdawi, S. et al. (2008). The Friedreich ataxia GAA repeat expansion
mutation induces comparable epigenetic changes in human and transgenic
mouse brain and heart tissues. Hum. Mol. Genet. 17, 735-746.
•Richards, R.I. (2001). Dynamic mutations: a decade of unstable expanded
repeats in human genetic disease. Human Molecular Genetics 10: 2187-
2194.
•Pearson, C. E., Edamura, K. N., Cleary, J. D. (2005). Repeat instability:
mechanisms of dynamic mutations. Nat. Rev. Gen. 6: 729-742.
Thank You

for Your Attention!

 Expansion mechanisms
Recombination
Replication Repair associated associated
associated instability
instability