Presentation Overview

I. Genomic Imprinting

II. Prader – Willi Syndrome (PWS)

III. Angelman Sydnrome (AS)

I. Genomic Imprinting
1. Definition
2. How it was discovered?
3. What is the purpose of this process?
4. Mechanism of genomic Imprinting
5. List of imprinted genes
6. What can effect to genomic imprinting?
7. Comparison between genomic imprinting and X –
inactivation
8. Diseases associated with genomic imprinting
1. Definition

 Two copies of each chromosome — and therefore, two

copies of each gene — arrive in every fertilized egg.
However, in some cases, one of the genes within a pair is
somehow switched "off" due to epigentically marking. This
process is known as genomic imprinting.

 Whether or not particular genes are inactivated in this way

depends on which sexes of parent the genes were inherited
from. In such cases, a gene is only expressed when it comes
from the "correct" parent (either mother or father).
Normally expressed gene
in both paternal and
maternal 15

Loci normally imprinted in chromosome 15

2. How it was discovered?

• Genomic imprinting was discovered almost simultaneously

by two laboratories in 1984. Prior to the publications of
McGrath and Solter (1984) and Surani et al. (1984),
conflicting results existed about whether both parental
genomes were required for normal development.
Illustration of Surani et al ‘s experiment in 1984
 Number of
Number of
Genetic Component Total
Viable Embryos
Pregnancies
Activated oocyte +
48 0
egg pronucleus
Activated oocyte +
24 9
sperm pronucleus

Summary of results from experiments examining the differential

role of egg or sperm pronuclei in mouse development
 Conclusion

• Maternal and paternal genomes are both essential for

the development of mice past early embryonic stages and
that their contribution to growth of the early conceptus are
not equivalent:

 Paternal genes are essential for growth of trophoblastic.

 Maternal genes can provide all of the essential of the

tissues of the embryo proper but lack essential activities
for growth of the trophoblast.
 Surani et al. speculated that during production of eggs
or sperm, some of the cells' genes are imprinted with an
indication of the gamete in which the genes began.
 First paternally
imprinted gene: IgFII

1991

First matermally
imprinted genes:
(Igfr2, H19)
3. What is the purpose of this?
1) The evolvability hypothesis

• Content:
 Increased evolvability on a population by masking a locus.
 The masked alleles can accumulate many mutations. This
is proposed to increase the rate of adaptive evolution
because it increases the probability of adaptive changes.

• Disadvantages of the hypothesis:

 Too general
 The reality is only a few hundred loci have been
discovered in humans that exhibit genomic imprinting, it is
simply too rare a genetic phenomenon to increase evolvability!
2. The ovarian time bomb hypothesis

• Content:
 The genes that are responsible for trophoblast development
are inactivated in the oocytes to prevent ovarian
trophoblastic disease. The active copies of these genes,
which are necessary for successful implantation, are then
provided by the sperm genome after fertilization.

• Disadvantages of the hypothesis:

 Too specific.
 Loci which have no relationship to oocyte development
imprinted
4. Mechanism of genomic imprinting

4.1 Requiremnts for any biochemical modification of the

DNA and/or chromatin which can account for imprinting

1)The modification must be made before fertilization.

2)It must be able to confer transcriptional silencing.
3)It must be stably transmitted through mitosis in somatic cells.
4)It must be reversible on passage through the opposite parental
germline.
4.2 Some fairly exotic possibilities that are consistent
with these requirements

 Specific DNA sequence.

 Oocyte or spermatocyte-specific DNA binding proteins.
 DNA methylation.
4.3 DNA methylation - the main and classical mechanism
for genomic imprinting

DNA Methylation
DNA Methylation
Affect of DNA methylation on transcription
 A new study has suggested a novel inheritable imprinting
mechanism in humans that would be specific
of placental tissue and that is independent of DNA
methylation.
4.4 Imprinting in cells

 In gametogenesis:
• An imprint can be erased and then reestablished
during gametogenesis to match the sex of the gamete-
producing parent.

 In embyro/offspring development:
• The epigenetic signal must be consistent in the
mature gamete and reversible in the next generation,
depending on the sex of the offspring.
Illustration of erasure and restablishment process of imprinted gene
5. Listed of imprinted gene

 As of 2014, there are about 150 imprinted genes

known in the mouse and about half that in humans
6. What can affect to genomic imprinting?

 Imprinted genes are especially sensitive to

environmental signals. Because imprinted genes have
only a single active copy and no back-up, any epigenetic
changes or "epimutations" will have a greater impact on
gene expression.

 Environmental signals can also affect the imprinting

process itself: diet, hormones and toxins can all affect
this process, impacting the expression of genes in the next
generation.
7. Comparison between genomic imprinting and
X - inactivation
7.1 Similaritites:

 Not mutation
 Due to epigenetics mechanisms such as DNA methylation.
 It is incomplete
 Inactivation is permanent in somatic cells and reversible in
developing germ line cells
 All imprinted genes are active during spermaogenesis /
oogenesis
 Essential for normal development of individuals.
7.2 Differences:

Genomic Imprinting X – inactivation

Happen in both sexes Only happen in female cells

Depend on the sexes of the Depend on the sexes of the

parents / offsprings offsprings only

Imprinting involves of genes Randomly inactive between two

contributed by a specific parent. X chromosomes

Only nine chromosomes are

50% of X chromosomes will be
known to have regions of genes
inactivated
that are imprinted
8. Diseases associated with genomic imprinting

 Improper imprinting or proper imprinting – linked

mutation can result in an individual having two active
copies or two inactive copies. This can lead to severe
developmental abnormalities, cancer, and other problems:

1) Prader Willi Syndrome

2) Angelman Syndrome
3) Silver-Russell syndrome
4) Beckwith-Wiedemann syndrome
5) Albright hereditary osteodystrophy
6) Uniparental disomy 14
II. Prader Willi Syndrome (PWS)
1. Overview
2. History
3. Signs and Symptoms
4. Genetics changes
5. Inheritance Pattern
6. Pathophysiology
7. Diagnosis – Prevention – Treatment
8. Epidemiology
1. Overview

• Prader-Willi syndrome is caused by the loss of function of

critical genes in a particular region of paternal
chromosome 15 while the genes in the maternal ones are
inactive due to genomic imprinting. These genes are vital for
the normal development of the nervous system.
Therefore, this loss may impair the neurons and contribute
severe problems such as intellectual disability, behavioral
problems, and the physical features of the disorder.
2. History

Andrea Alexis Heinrich

Prader Labhart Willi
2. History

• First report the pattern of abnormalities that

are now known to be symptoms of the
syndrome was in 1956.

• The 1980s and 1990s saw the confirmation

of the genetic cause of the syndrome and
the development of genetic tests for Prader-
Willi syndrome and the genetic subtypes.
3. Signs and Symptoms

• Poor muscle tone (hypotonia)

• A lack of eye coordination (Strabismus)

• Slow development, intellectual impairment

• Abnormal neurologic function

• Feeding difficulties (due to poor muscle tone)

• Speech and behavioural problems, sleep disorders

• Complications: obesity, type 2 diabetes

Facial features of PWS patients
4. Genetics changes
4.1 What genes are lost?

• Prader-Willi syndrome is caused by the loss of function

of genes in 15q11-13 region of paternal chromosome 15.

Regions in Chromosome 15 (marked with red) which caused

Prader – Willi Syndrome
4. Genetics changes
4.1 What genes are lost?
This leads to the missing of the copies of these genes:

 SNRPN
 Necdin (NDN)
 Clusters of snoRNAs: SNORD64, SNORD107, SNORD108
 2 copies of SNORD109
 29 copies of SNORD116 (HBII-85)
 48 copies of SNORD115 (HBII-52)
4. Genetics changes
4.1 What genes are lost / defect?

 Normally, the maternally inherited copies of these

genes are virtually silent, only the paternal copies of
the genes are expressed (monoallelic expression).
Thus, if this region in paternal chromosome is lost or
defect, there are not any expression of these genes
anymore.

Prader – Willi Syndrome

4.2 Abnormalities of chromosome that lead the loss
of function of genes in 15q11-13 region
• Type of abnormalities: Deletion of 15q11 – 13 region
• Frequency: about 70%
• Consequence: missing critical gene
4.2 Abnormalities of chromosome that lead the loss
of function of genes in 15q11-13 region

• Type of abnormalities: Maternal uparental disomy(UPD)

• Frequency: about 25%
• Consequence: no working copies of certain genes
4.2 Abnormalities of chromosome that lead the loss
of function of genes in 15q11-13 region

• Type of abnormalities: Imprinting defect

• Frequency: less then 5%
• Consequence: PWS section may not be switched on correctly
4.2 Abnormalities of chromosome that lead the loss
of function of genes in 15q11-13 region

• Type of abnormalities: Chromosome translocation

• Frequency: very rare
• Consequence: unbalanced gametes which lost partenal
15q11-13 region
5. Inheritance pattern

• Most cases of Prader-Willi syndrome are not inherited,

particularly those caused by a deletion in the
paternal chromosome 15 or by UPD.

• Affected people typically have no history of the disorder in

their family.

• Rarely, a genetic change responsible for Prader-Willi

syndrome can be inherited.
6. Pathophysiology

The exact mechanisms of

PWS are unclear
6. Pathophysiology

• Although three paternally expressed gene have been

identified within the deletion region, no correlation
between the loss of expression of a specific PWS
region gene and the PWS phenotype has been shown.

• No affected individuals have been identified in whom a

mutation affects the expression only one PWS region gene,
suggesting that PWS is a contiguous gene syndrome in
which the loss of expression of several genes is
required for full manifestation of the disorder

MacDonald HR, Wevrick R (1997)

 NDN gene
SNPRN snoRNAs
gene gene

Prader- Willi
Syndrome
6.1 Small nuclear ribonucleoprotein Polypeptide N(SNPRN)

 Function of SNPRN: pre-mRNA processing.

 Hypothesis about the role of SNPRN in the

development of PWS: (Reed ML, Leff SE -1994)

• Sequence polymorphism has shown that human SNRPN

is monoallelically expressed in fetal brain and heart and
in adult brain.
• Analysis of maternal DNA and SNRPN cDNA confirmed
that the maternal allele of SNRPN is not expressed in fetal
brain and heart.
 Paternal absence of SNRPN is responsible for the
PWS phenotype.
 Expression studies and the role of SNPRN in PWS:
Molecular analysis of PWS by RT-PCR of SNRPN expression
6.2 Necdin (NDN) gene

• Function of NDN: regulate the permanent arrest of cell

growth of post – mitotic neurons during development.

• Hypothesis about the role of NDN in the development of

PWS: necdin deficiency in individuals with PWS may cause
part or all of their neurological deficit by interference with
normal brain development.
Melanoma-Associated Antigen Domain

Necdin’s domain
Necdin interacts with other proteins to
regulate the differentiation of cells
 Expression studies and the role of NDN in PWS:
6.3 Small nucleolar RNA (snoRNAs)

• Function of snoRNAs: guide chemical modifications of

other RNAs, mainly ribosomal RNAs, transfer RNAs and small
nuclear RNAs.
 Hypothesis about the role of snoRNA in the
development of PWS:

• Loss of a particular group of snoRNA genes, known as the

SNORD116 cluster, may play a major role in causing the
signs and symptoms of Prader-Willi syndrome.

• Studies of human and mouse model systems have

deletions of 29 copies of SNORD116 (HBII-85) to be the
primary cause of Prader–Willi syndrome.
Role of snoRNA in the development of PWS
Expression analysis of SNORD115 and SNORD116 among 20
different human tissues
7. Diagnosis
7.1 Diagnosis– Prevention - Treatment

• Clinical presentation
 Facial features of Prader – Willi
Syndrome
 Libido Impairment
 Undescended testicle
 Hypotonia
 Itellectual impairment
 Obesity
Some common symptoms are used to diagnose PWS
(Studies from Central Pediatric Hospital, Ha Noi Medical School)
• Genetic testing

 The mainstay of diagnosis is genetic testing, specifically

DNA-based methylation testing to detect the absence of the
paternally contributed Prader–Willi syndrome region
on chromosome 15q11-q13.

 Such testing detects over 97% of cases.

7.2 Prevention
• There is currently no known method for preventing
Prader-Willi syndrome.
• Genetic testing and counseling.
• Preventing complications of obesity.
7.3 Treatment

There is no cure for

Prader – Willi Syndrome
7.3 Treatment

 Treatment is to lessen the symptoms, including:

• Speech therapy

• Physiotherapy during infancy

• Growth hormone therapy

• Hormone therapy with sex hormones

7.3 Treatment
 Target therapy:

 Research is focusing on targeting specific genes for

treatment. Current treatment focuses on managing the
medical and developmental issues.
8. Epidemiology

 Prader-Willi syndrome affects an estimated 1 in 10,000

to 30,000 people worldwide.

(US National Library of Medicine)

III. Angelman Syndrome (AS)
1. Overview
2. History
3. Signs and Symptoms
4. Genetics changes
5. Inheritance Pattern
6. Pathophysiology
7. Diagnosis – Prevention – Treatment
8. Epidemiology
1. Overview

• Angelman syndrome is caused by the loss of function of

UBE3A in a particular region of maternal chromosome
15 while the gene in the paternal ones is inactive due to
genomic imprinting. This gene encodes for Ubiqutin Ligase
E3 (UEB3A) which plays a critical role in ubiqutination
pathways in the nervous system. Hence, when UBE3A
protein is no longer functioning properly, it may contribute to
delayed development, intellectual disability, severe speech
impairment, and problems with movement and balance
(ataxia).
2. History

• Harry Angelman, a pediatrician working in Warrington,

England, first reported three children with this condition
in 1965.
"Boy with a Puppet" or "A child
with a drawing" by Giovanni
Francesco Caroto.
• Case reports from the United States first began
appearing in the medical literature in the early 1980s.

• In 1987, it was first noted that around half of the

children with AS have a small piece of chromosome
15 missing (chromosome 15q partial deletion).
3. Sign and Symptoms

• Happy, excitable personality

• Developmental delays, including no crawling or

babbling at 6 to 12 months

• Intellectual disability

• No speech or minimal speech

• Difficulty walking, moving or balancing well (ataxia)

• Frequent smiling and laughter

3. Sign and Symptoms

 Others complications can have:

• Seizures, usually beginning between 2 and 3 years of age.

• Stiff or jerky movements.
• Small head size, with flatness in the back of the head
(microbrachycephaly).
• Tongue thrusting.
• Hair, skin and eyes that are light in color (hypopigmentation).
• Unusual behaviors, such as hand flapping and arms uplifted while
walking.
4. Genetics changes
4.1 What genes are lost / defect?
• Angelman syndrome is caused by the loss of function of
UBE3A gene in 15q11-13 region of maternal chromosome 15.

Regions in Chromosome 15 which caused Angelman Syndrome

4. Genetics changes
4.1 What genes are lost / defect?

•Both copies of this gene are turned on (active) in many

of the body's tissues.

•In certain areas of the brain, however, only the copy

inherited from a person's mother (the maternal copy) is
active.

•If the maternal copy of the UBE3A gene is lost because of a

chromosomal change or a gene mutation, a person will have
no active copies of the gene in some parts of the brain

Angelman Syndrome
4.2 Abnormalities of chromosome that lead the loss
of function of gene in 15q11-13 region
• Type of abnormalities: Deletion of 15q11 – 13 region
• Frequency: about 70%
• Consequence: missing UBE3A gene
4.2 Abnormalities of chromosome that lead the loss
of function of gene in 15q11-13 region

• Type of abnormalities: Imprinting Center Defect

• Frequency: about 6%
• Consequence: inactivates UBE3A gene
4.2 Abnormalities of chromosome that lead the loss
of function of gene in 15q11-13 region

• Type of abnormalities: UBE3A mutation

• Frequency: about 11%
• Consequence: prevent its expression or function normally
4.2 Abnormalities of chromosome that lead the loss
of function of gene in 15q11-13 region

• Type of abnormalities: Paternl Uniparental Disomy (UPD)

• Frequency: about 3%
• Consequence: no working copies of UBE3A
4.2 Abnormalities of chromosome that lead the loss
of function of gene in 15q11-13 region
• Type of abnormalities: Chromosome translocation
• Frequency: very rare
• Consequence: unbalanced gametes which lost martenal
15q11-13 region
5. Inheritance pattern

• Most cases of Angelman syndrome are not inherited,

particularly those caused by a deletion in the
maternal chromosome 15 or by paternal uniparental disomy.

• Affected people typically have no history of the disorder in

their family.

• Rarely, a genetic change responsible for Angelman

syndrome can be inherited.
6. Pathophysiology

UBE3A

Ubiquitin Ubiquitination

Angelman
Syndrome
6. Pathophysiology
6.1 Structure and function of Ubiqutin

 Structure of Ubiquitin:

• A small protein that exists in all eukaryotic cells.

• The ubiquitin protein itself consists of 76 amino acids
• Molecular mass of is about 8.5 kDa.
• Key features include: C-terminal tail and 7 lysine
residues
• It is highly conserved among eukaryotic species: Human
and yeast ubiquitin share 96% sequence identity
6. Pathophysiology
6.1 Structure and function of Ubiqutin

Cartoon view of ubiquitin protein

6. Pathophysiology
6.1 Structure and function of Ubiqutin

 Function of Ubiquitin:

• Degradation of the protein via the proteasome

• Apoptosis (cell death)
• DNA transcription and repair
• Organelle biogenesis
• Processing of antigens
• Viral infection
Degradation of Misfolded Proteins By Proteasomes
Proteins marked by a polyubiquitin chain are degraded by the
proteasome
6. Pathophysiology
6.2 Structure and function of Ubiquitin protein ligase E3A
(UBE3A)

 Structure of UBE3A:

• Dry – weight: 142510.89 dalton, including 9681 atoms.

• It has in total 4 chains represented by 2 sequence-unique.

• Possess one of two domains: the homologous to the E6-AP

carboxyl terminus (HECT) domain and the really interesting
new gene (RING) domain (or the closely related U-box
domain)
6. Pathophysiology
6.2 Structure and function of Ubiquitin protein ligase E3A
(UBE3A)

Cartoon view of Ubiquitin protein ligase E3A

6. Pathophysiology
6.2 Structure and function of Ubiquitin protein ligase E3A
(UBE3A)

 Function of Ubiquitin: E3 ubiquitin ligases catalyse the

final step of the ubiquitination cascade that liagates
ubiquitins to the target protein.
6. Pathophysiology
6.3 Ubiquitination – Ubiquitin Pathway

Key features:

• Enzymatic post-translational modification in which a

ubiquitin protein is attached to a substrate protein.

• Requires three types of enzyme: ubiquitin-activating

enzymes, ubiquitin-conjugating enzymes, and ubiquitin
ligases, known as E1s, E2s, and E3s, respectively

• Cosists of three 3 steps: activation, conjugation, ligation.

6. Pathophysiology
6.3 Ubiquitination – Ubiquitin Pathway

We would like invite you all to watch this video!

6. Pathophysiology
6.3 The role of Ubiquitin protein ligase E3A in the
normal development and function of the nervous system

• Controls (regulate) the balance of protein synthesis and

degradation (proteostasis) at the junctions between nerve
cells (synapses) where cell-to-cell communication takes
place.

• Regulation of proteostasis is important for the synapses to

change and adapt over time in response to experience, a
characteristic called synaptic plasticity. Synaptic plasticity is
critical for learning and memory.
Gene expression in eukaryotic cells can be
controlled at various steps
7.1Diagnosis
7. – Prevention - Treatment
Diagnosis:

• A blood test: can detect up to 80-85% of individuals with

Angelman syndrome by identifying whether the UBE3A
gene is functioning properly.
• Clinical diagnosis: For the remaining 15-20% of
individuals, an experienced clinician who is familiar with
Angelman syndrome.
 A combination of genetic tests can reveal the
chromosome defects related to Angelman syndrome. These
tests may review:

• Parental DNA pattern: This test, known as a DNA

methylation test, screens for three of the four known
genetic abnormalities that cause Angelman syndrome.

• Missing chromosomes: A chromosomal microarray

(CMA) can show if portions of chromosomes are missing.

• Gene mutation: Rarely, Angelman syndrome may occur

when a person's maternal copy of the UBE3A gene is active,
but mutated. If results from a DNA methylation test are
normal, your child's doctor may order a UBE3A gene
sequencing test to look for a maternal mutation.
7.2 Prevention:

• Because most cases of Angelman syndrome are caused by

spontaneous genetic mutations, there is currently no
known way to prevent the disease.
• Genetic counseling: to assess risks to siblings and other
family members is based on knowing the mechanism
involved in causing the loss of expression of this
genetic region at the molecular level.
7.3 Treatment:

There is no cure for

Angelman Syndrome
 Depending on patients’s signs and symptoms,
treatment for Angelman syndrome may involve:

• Anti-seizure medication
• Physical therapy
• Communication therapy
• Behavior therapy
 Target therapy:

 Research is focusing on targeting specific genes for

treatment. Current treatment focuses on managing the
medical and developmental issues.
8. Epidemiology

 Angelman syndrome affects an estimated 1 in 12,000

to 20,000 people
(US National Library of Medicine)
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