complex analysis best book for solving questions.pdf
Imprinting.ppt
1. Genomic Imprinting
•Definition- Differential expression of
two parental alleles
• Only occurs in eutherians (placental,
nonmarsupial) mammals
• Not in other vertebrates
•Of 20-some identified genes, most are
involved in
1. Fetal growth
• Igf2, IgF2r, H19, Grb1
2. Brain development
• Prader-Willi syndrome (PS), Angelman syndromes (AS),
Peg1/Mest
2.
3.
4.
5.
6. Genomic-imprinting conflict
What is genomic imprinting and why has it evolved?
Expression of a gene depending on whether inherited
from father or mother
Main arenas of imprinting effects on human health:
-Placenta
-Brain
-Carcinogenesis
-Stem cells
-In vitro fertilization
from dad from mom
7. • In experiments on mice
maternal and
paternal
pronuclei
normal mouse
both
paternal
pronuclei
The placenta is
huge, the embryo
is undeveloped
both
matern
al
pronucl
ei
The embryo is
formed, the placenta
is underdeveloped
9. CONFLICT THEORY OF IMPRINTING
-> abundant support from empirical studies of imprinted
genes and growth, in mice and humans
(1) IGF2-IGF2R (Haig & Graham 1991 Cell)
(2) CDKN1C (Andrews et al. 2007 BMC Dev Biol)
(3) GRB10 (Charalambous et al. 2003 PNAS)
Beckwith-
Wiedemann
Syndrome
Silver-
Russell
syndrome
Mighty mouse
Normal sized
human
2 doses IGF2
1 dose IGF2
0 doses IGF2
10. Genomic imprinting
the unequal expression of the maternal and paternal alleles of a gene
XX
X
Y
Maternal allele
Patternal allele
Patternal
allele
Matternal
allele
11. • Two major clusters of imprinted genes have been identified in
humans, one on the short (p) arm of chromosome 11 (at
position 11p15) and another on the long (q) arm of chromosome
15 (in the region 15q11 to 15q13).
20. Paternally expressed imprinted genes that function as growth
promoters (i.e., Igf2, Peg1, Peg3, Rasgrf1, Dlk1) and show
growth retardation in embryos deficient for the gene. There
are also maternally expressed imprinted genes that function
as growth repressors (i.e., Igf2r, Gnas, Cdkn1c, H19, Grb10),
as shown by a growth enhancement in embryos deficient for
the gene.
21. Categories of imprinted genes
1. Fetal growth genes- Insulin-like growth
factor-like II (IGF2) response pathway
• IGF2
• Igf2r
• Grb10
• H19
• Gnas
• Rasgrf
• Mash2
• Why?- Embryo develops in a parasite-like
relationship with mother.
23. Prader-Willi Syndrome
• 1 in 15,000 live births
• mostly sporatic
• deletion at 15 q11-q13
• diagnosis at 2 years
• compulsive overeaters
24. A typical Prader-Willi patient
Prader Willi Syndrome- Due to
paternal chromosome deletion
25. • 1 in 25,000 live births
• mostly sporatic
• 80 % have deletion at 15q11-q13
• Specifically mutation of UBE3A gene
Angelman Syndrome
26. Angelman Syndrome
•Speech impairment
•None or minimal use of words
•Receptive and non-verbal communication skills higher than
verbal ones
•Movement or balance disorder, usually ataxia of gait
•Behavioral uniqueness: any combination of frequent
laughter/smiling; apparent happy demeanor
•easily excitable personality, often with hand flapping
movements; hypermotoric behavior; short attention span
28. Parent Offspring Conflict Hypothesis (Haig
hypothesis)
• Conflict between male and female over allocation
of maternal resources to offspring
• Dad uses imprinting to direct all resources to
immediate offspring (not future litters)
• Mom uses imprinting to allocate resources to
multiple litters
• Thus, predict paternally expressed genes would
promote growth, maternally expressed genes should
slow it down
• Prediction mostly hold true
• Example- Igf2 (paternally expressed)-if defective=40%
reduction in growth
29. Parent Offspring Conflict Hypothesis (Haig
hypothesis)
Example – The Igf2 gene and its receptor Igf2r
•Igf2 (paternally expressed)-if defective=40%
reduction in growth
•Igf2r (Igf2 receptor)- if defective=increase growth
•Igf2-/Igf2r- = normal
Another test- Ask if imprinting fails to occur in a monogomous
species
The Beach mouse is entirely monogomous
….but imprinting still occurs, contrary to model
31. How are imprinted genes silenced?
S. Tilghman, Cell 96:185
Dnmt-/- mice-
Many imprinted genes
(e.g. H19) reactivated
..but, Igf2 and Igf2r are
silenced
Mechanism- Methylation interferes with transcription factor binding
Problems with model-
1. Promoters of silent Igf2 and Igf2r alleles are unmethylated
2. One gene, Mash2, is unaffected by loss of methylation
32. How are imprinted genes silenced?
S. Tilghman, Cell 96:185
Mechanism-
Promoters compete
for a single
enhancer
Problem with model-
Both H19 and Igf2 are expressed if H19 gene replaced
with luciferase
Igf2 H19
33. How are imprinted genes silenced?
S. Tilghman, Cell 96:185
Epigenetic marker binds to
unmethylated DNA
Mechanism-
Methylation serves two
purposes
1. Inactivate a gene (e.g.
H19)
2. Prevent binding of
epigenetic marker so
that Igf2 is activated
Igf2 H19
Epigenetic insulator prevents enhancer from “talking to” Igf2
Evidence in support: if delete insulator element- both Igf2 and H19 expressed
34. Evidence for chromatin boundary mechanism
Deletion of ICR- both genes expressed
Identify protein (called CTCF) that binds ICR
CTCF cannot bind methylated DNA
Thorvaldsen and Bartolmei, Science 288:2145, 2000
35. How are imprinted genes silenced?
S. Tilghman, Cell 96:185
Mechanism- Antisense
transcription of
unmethylated
chromosome blocks
sense strand transcription
Mechanism- Antisense
RNA blocks sense strand
transcription