International University, Vietnam National University – HCMC

Genetics

Assignment 2:

Course name: Genetic

Instructor: Tong Thi Hang

Group members: Vu Thi My Huyen – BTBTIU17135

Dang Gia Hoang – BTBTIU17071

Nguyen Ngoc Thien Kim – BTBTIU17093

Section: Wednesday Morning Date: 14/06/2019

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 International University, Vietnam National University – HCMC
Genetics

TABLE OF CONTENT
❖ Introduction: ...................................................................................................................................... 3
❖ Definition ............................................................................................................................................ 3
• Trinucleotide repeat expansion: .............................................................................................. 3
• Microsatellites: ............................................................................................................................. 3
❖ General mechanism and why it causes diseases in human: ............................................... 4
6 common factors contributed to pathogenic mechanism of repeat expansion diseases .. 6
1. Sequence of repeat ...................................................................................................................... 6
2. Size of repeat................................................................................................................................. 6
3. Location of within gene .............................................................................................................. 7
4. Whether repeat encodes RNA or protein ............................................................................... 7
5. Function of repeat - containing gene...................................................................................... 7
6. Extent of meiotic and somatic instability .............................................................................. 7
❖ Examples of toxic gain of functions diseases: ........................................................................ 8
• CAG/ poly Q disease: .................................................................................................................. 8
• Huntington’s disease like 2: ...................................................................................................... 9
• DM1,2: ........................................................................................................................................... 10
• FXTAS: .......................................................................................................................................... 12
❖ Examples of loss of functions diseases .................................................................................. 14
• Friedreich's ataxia...................................................................................................................... 14
• Fragile X syndrome: .................................................................................................................. 15
❖ Conclusion: ..................................................................................................................................... 16
❖ References:...................................................................................................................................... 17

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Genetics

❖ Introduction:
• When mentions the disease caused by genetic disorders, there are
many reasons for that. E.g. mutation in one gene (monogenic
disorder), by mutations in multiple genes (multifactorial inheritance
disorder), by a combination of gene mutations and environmental
factors, by damage to chromosomes (changes in the number or
structure of entire chromosomes, the structures that carry genes) or
by slippage strand occurred in DNA replication, the crossing over
unequal and ethical can cause the expansion repeat nucleotide

• One kind of genetic disorders are the nucleotide repeat extensions

which caused diseases in human. The more nucleotide repeats
extensions, the more disease severity. Researching the complex
mechanism that nucleotide repeat extensions cause diseases in
human is necessary.

❖ Definition
• Trinucleotide repeat expansion:
o A gene has a triplet repeat expansion encoded the protein means
the increase of the triplet (trinucleotide) repeats in the gene
sequence.
o The difference between each triplet repeat expansion is the length, it
is usually polymorphic. The more repeat length, the more disease
severity.

• Microsatellites:
o In noncoding regions between genes or intron has the repetitive
segments of DNA dispersive in the genome. Since their naturally
occurring high variability in repeat number between individuals, they
are usually used as markers for linkage analysis. These regions are
inherently genetically unstable and susceptible to mutations.

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Genetics

❖ General mechanism and why it causes diseases in human:

• Nucleotide repeat expansion generally effects on the process from
DNA to protein through transcription and translation.

• Depending on the location or length of nucleotide repeat (coding or

non-coding region...), the level of gene expression and type of
diseases are performed such as loss or gain of function (proteins are
no longer have correctly-working function) or toxic mRNA (having
negative effects to other mRNA or proteins).

• There are two main types of triplet repeats diseases.

o The first type is that the expanded triplet repeat disorders is

translated, usually including poly-glutamine or poly-alanine homo-
repeats.

o In the second type, triplet repeats which are present in non-coding

regions of the gene can be expressed at the mRNA level (affecting
expression levels of coded proteins) but are not translated, so it have
no effect on protein structure. However, the produced mRNA
sometimes give rise to highly repetitive peptides of different length
and amino acid content by translating. The formation of a toxic
homo-polypeptide might be derived from this step which can lead to
the development of the pathology

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Genetics

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 International University, Vietnam National University – HCMC
Genetics

6 common factors contributed to pathogenic mechanism

of repeat expansion diseases

1. Sequence of repeat
• Disease cause by repeat expansion nucleotide depend on which kind
of repeat sequence. For examples, repeat of CAG triplet nucleotide
affect to gene expression. That means its make mistake in
transcription, translation, and protein modification process.
• As the result, encoding disease protein cause at least nine disease
related to neurodegeneration.
2. Size of repeat
• Different expanding nucleotide repeats range cause to several
genetic diseases.
• For instance, Fragile – X syndrome disease because of CGG triplet
nucleotides repeat from 50 to 1500 times. While, with the same
repeat sequence CAG, but the repeat copies sequence from 100 to
1000 cause to Jacobsen syndrome disease.

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Genetics

3. Location of within gene

• Repeat sequence can occur in non-coding or coding regions of
genes. In Huntington disease, trinucleotide CAG expands in the
coding region make the abnormal long strand of glutamine lead to
dominant inherited gain-of-function mutation.
• Whereas, in case of DM1 disease cause by dominant inherited non-
coding repeat expansions. So, it elicits toxicity through gain-of-
function mechanism as RNA.
4. Whether repeat encodes RNA or protein
• Fragile X tremor ataxia syndrome cause by a CGG triplet repeat
expansion within 5’ UTR of the Fragile X gene. That cause RNA
toxicity, which results in the binding of the CGG-binding protein.
• So, it prevents mRNA transcription and splicing, as well as dendritic
mRNA transport.
• In addition, trinucleotides CGG expansion induce RAN translation
(non- AUG translation) within 5’ UTR of Fragile X gene through AUG
independent mechanism, which generates a polyglycine peptide toxic
to cells.
5. Function of repeat - containing gene
• Expansion repeat nucleotide within gene can cause loss-of-function
or gain-of-function. That cause gene regulation different and can
cause bad or good in human as well.
• For examples, Friedreich Ataxia expand GAA triplet located in intron
region (non-coding region) cause transcription client or down-
regulation of the associated gene. Hence, its act as recessive
inherited, loss-of-function mutation.
6. Extent of meiotic and somatic instability
• Huntington disease has variation in onset age but with an average
age at onset of 40 years. The length of CAG repeat mutation primarily
determined the age onset of Huntington disease.
• The Huntington disease is somatic instability, underlying progressive
length increase, particularly in brain regions that are targets of
neurodegeneration. Moreover, the longer the repetition, the greater

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Genetics

the likelihood of somatic instability that gains associated with earlier

disease onset.
❖ Examples of toxic gain of functions diseases:
• CAG/ poly Q disease:
o The polyglutamine disease have nine common diseases. Nine types
of polyglutamine diseases are dominant inherited disorders except
for SMBA which an X-linked dominant toxic disorder feature is. That
have similar progressive and are major cause neurodegenerative
disease. That include Huntington's disease, Spinal and Bulbar
Muscular Atrophy ( SMBA), Dentatorubral-pallidoluysian atrophy (
DRPLA) , and the Spinocerebellar ataxia( SCA) types 1,2,3,6,7, and
17. The disease cause by the expansion of unstable CAG triplet
nucleotide in the gen-coding region. As the result, glutamine
stretching in the protein, which makes host protein toxic through
gain-of-function mechanism. So, triple nucleotide CAG repeat
influence encode disease protein and make several diseases
related to neurodegeneration.

o Moreover, each disease has different wildtypes, so the range of

repeating make disease also different as well as different affect in
human health

o Mechanism of CAG/ poly Q disease:

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Genetics

• Huntington’s disease like 2:

o Huntington disease like 2 is caused by a CTG repeat expansion in
the Junctophilin 3 (JPH3) from 41 to 58 times. This mutation
increasing the length of DNA, which cause to the production of an
abnormal JPH3. The JPH3 genes contribute to making proteins that
important for brain function. Therefore, the encoding disease protein
build-up in nervous cells and because cells function abnormal and
cause several disease and symptom as same as Huntington disease.

o Mechanism of Huntington’s disease like 2:

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Genetics

• DM1,2:
o Myotonic Dystrophy or DM is a dominantly complex inherited genetic
disorder that is the most common cause of muscular dystrophy in
adults.

o There are 2 different forms of diseases which are caused by different

microsatellite expansions in 2 loci:

o Myotonic dystrophy type 1 or DM is caused on chromosome 19 which

contain an expansion of a CTG repeat located in the 3' untranslated
region of DMPK (myotonic dystrophy protein kinase)

o Myotonic dystrophy type 2 or DM2 is caused by an unstable CCTG

repeat in intron 1 of ZNF9 (zinc finger protein 9) on chromosome 3.

o And both of similar features that DM1 and DM2 having is that they
are caused by a repeat expansion in a region transcribed into RNA
but not translated into protein. The mutant RNA transcripts of DM1
and DM2 aberrantly affect the splicing of the same target RNAs, such
as chloride channel 1 (ClC-1) and insulin receptor (INSR), resulting in
their shared myotonia and insulin resistance.

o The pathogenic mechanism of Myotonic Dystrophy involves the RNA

transcribed from the expanded allele containing long tracts of
(CUG)(n) or (CCUG)(n). The toxic effect of RNA through two RNA-
binding proteins is MBNL1 (muscle blind-like 1) and CUGBP1 (CUG-
binding protein 1) which is relevant to other RNA dominant disorders.

o In specific, RNA transcripts likely form double-stranded hairpin

structures from the extended tracts of CUG or CCUG repeats, to
which MBNL proteins prefer to bind. MBNL proteins co-localize with
the ribonuclear inclusions formed by mutant RNA but the nuclear
sequestration of MBNL which directly leads to a significant decrease
in normal MBNL activity, or whether expanded CUG/CCUG RNA
signals an alternate pathway to MBNL functional inhibition is unclear.
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Genetics

CUG-BP1 levels are increased in DM1 cells, independent of MBNL

protein regulation and increased CUG-BP activity and/or loss of
MBNL function may lead to aberrant gene splicing events associated
with DM1 and DM2 as shown.

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Genetics

• FXTAS:
o Fragile X-associated tremor/ataxia syndrome (FXTAS) which are
caused by a CGG triplet repeat expansion within the 5′ UTR of FMR1
is a neurodegenerative disorder. Normally, individuals who possess
between 5 and 54 CGG repeats, and full mutation CGG repeats
greater than 200 can have the neurodevelopmental disease fragile X
syndrome (FXS). Therefore, the FXS lead the owner to the excessive
methylation of FMR1 and loss of FMRP protein. Moreover, the
individuals with 55–200 CGG repeats are referred to as premutation
carriers.

o RNA toxicity and repeat associated non-AUG translation (RAN)

protein toxicity (via RAN) are two widely accepted mechanisms for
the pathogenesis of FXTAS.

o There are several evidences relate to the RNA toxicity mechanism.

o First, older adults do not develop FXTAS, who do not express FMR1
mRNA and lack FMRP, are full of the mutation (>200 repeats)

o Second, in FXTAS, formation of nuclear RNA aggregates since the

significant upregulation (2–8 fold) of the expanded CGG-repeat
FMR1 mRNA which bind with proteins to prevent them from
performing their normal biological functions, such as mRNA
transcription and splicing, as well as dendritic mRNA transport. The
level of FMR1 protein in cells from premutation carriers, however,
remains relatively unaltered.

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Genetics

o Third, besides the RNA-binding proteins (RBPs), these inclusions

which contain proteins do not bind to CGG-repeat on mRNA and are
reminiscent of the neuronal intranuclear inclusions founding in
protein-mediated neurodegenerative disorders and polyglutamine
diseases. Therefore, a protein-driven mechanism of FXTAS
pathogenesis was uncovered, in which the premutation CGG repeat
expansion was found to induce RAN translation within the 5′ UTR of
FMR1 mRNA via an AUG-independent mechanism. The resulting
polyglucan-containing protein, FMRpolyG, is present in the brains of
FXTAS patients and was found to be toxic to human cell lines.

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Genetics

❖ Examples of loss of functions diseases

• Friedreich's ataxia
o Friedreich's ataxia (FRDA) is a neurodegenerative disease caused by
reduced expression of the mitochondrial protein frataxin.

o Mechanism of Friedreich's ataxia:

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Genetics

• Fragile X syndrome:
o Fragile X syndrome affects a child's learning, behavior,
appearance, and health. Symptoms can be mild or more
severe. Boys often have a more serious form of it than girls.

o Mechanism of Fragile X syndrome:

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Genetics

❖ Conclusion:
In summary, extended repeat sequences cause more than 40
diseases most related to the nervous system. The disease is based
on two main mechanisms: gain-of-function mutations and loss-of-
function mutations. Based on repeated extension chains, scientist will
know more precisely the function of gene products, how to repeat
effects on gene expression. Moreover, the disease of repeat
expansion nucleotide provides a mechanism to cause disease. This
helps doctors to correctly diagnose the disease and find a way to
prevent and inhibit the disease. And, opens new ways of studying
pathogens that cause disease in humans.

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Genetics

❖ References:
▪ www.uniprot.org
▪ https://www.genome.gov/For-Patients-and-Families/Genetic-
Disorders
▪ https://www.cancer.gov/publications/dictionaries/genetics-
dictionary/def/microsatellite
▪ https://academic.oup.com/hmg/article/19/R1/R103/624761
▪ https://www.pulsus.com/scholarly-articles/trinucleotide-repeat-
diseases--antecipation-
diseases.pdf?fbclid=IwAR3otvAZx_15ObNazcLCHU7eXF6fvTk
63VyxzG5s9i5_qnh2RLHRdQ4K8HY
▪ https://www.ncbi.nlm.nih.gov/pubmed/16876389
▪ https://www.ncbi.nlm.nih.gov/pubmed/19909263
▪ https://www.sciencedirect.com/science/article/pii/S0925443906
000986?via%3Dihub
▪ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5418347/
▪ https://www.google.com/search?q=nucleotide+repeat+expansi
on+disease&rlz=1C1ASUM_enVN736VN736&source=lnms&tb
m=isch&sa=X&ved=0ahUKEwii_9yk8OLiAhUHc3AKHSppAr4
Q_AUIECgB&biw=1920&bih=925#imgrc=gA9IdAwh77Eq6M:
▪ Epigenetic Mechanisms in Repeat Expansion Disorders
▪ Fang He, Peter K. Todd
▪ Semen Neurol. Author manuscript; available in PMC 2013 May
16.
▪ Published in final edited form as: Semen Neurol. 2011 Nov;
31(5): 470–483. Published online 2012 Jan 21
▪ Fragile X-Associated Tremor/Ataxia Syndrome: From
Molecular Pathogenesis to Development of Therapeutics
▪ Ha Euna Kong, Juan Zhao, Shun Liang Xu, Peng Jinn, Yan
Jinn
▪ Front Cell Neurosis. 2017; 11: 128. Published online 2017 May
5
▪ Repeat expansion disease: Progress and puzzles in disease
pathogenesis

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Genetics

▪ Albert R. La Spada, J. Paul Taylor

▪ Nat Rev Genet. Author manuscript; available in PMC 2016 Jan
7.
▪ Published in final edited form as: Nat Rev Genet. 2010 Apr;
11(4): 247–258
▪ Huntington’s Disease Genetics
▪ Richard H. Myers
▪ NeuroRx. 2004 Apr; 1(2): 255–262
▪ Somatic expansion of the Huntington's disease CAG repeat in
the brain is associated with an earlier age of disease onset
▪ Meera Swami, Audrey E. Hendricks, Tammy Gillis, Tiffany
Masood, Jayalakshmi Mysuru, Richard H. Myers, Vanessa C.
Wheeler
▪ Hum Mol Genet. 2009 Aug 15; 18(16): 3039–3047. Published
online 2009 May 23. doe: 10.1093/mg/ddp242
▪ Aggregation Formation in the Polyglutamine Diseases:
Protection at a Cost?
Tiffany W. Todd, Janghoo Lim
Mol Cells. 2013 Sep 30; 36(3): 185–194. Published online
2013 Jun 19
▪ https://ghr.nlm.nih.gov/condition/huntington-disease-like-
syndrome?fbclid=IwAR1Z9ukH3M1rPpN_yVf4QFVpdq7kCNgF
l2EjQYYN9UnsiB5Dl7K7Ly56alI#sourcesforpage

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