Scribd Logo
Upload

Welcome to Scribd!

  • 31 views

    Genomic Imprinting & Epigenetics: 徐国良 研究员 细胞大楼 300 电话 E-mail: [email protected]

    Uploaded by

    api-3700537
    This document discusses epigenetic regulation of phenotypes through genomic imprinting and DNA methylation. It provides an overview of genomic imprinting, describing the discovery of imprinting and key examples like Igf2/H19. It also discusses DNA methylation, covering topics like the basic concepts, techniques for studying methylation, functions of methylation, mechanisms of methylation-mediated repression, and relationships between methylation and disease. Finally, it briefly introduces epigenetic regulation of phenotypes in Drosophila development and cancer through examples of homeotic gene regulation and implications for disease.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd

    Genomic Imprinting & Epigenetics: 徐国良 研究员 细胞大楼 300 电话 E-mail: [email protected]

    Uploaded by

    api-3700537
    31 views53 pages
    This document discusses epigenetic regulation of phenotypes through genomic imprinting and DNA methylation. It provides an overview of genomic imprinting, describing the discovery of imprinting and key examples like Igf2/H19. It also discusses DNA methylation, covering topics like the basic concepts, techniques for studying methylation, functions of methylation, mechanisms of methylation-mediated repression, and relationships between methylation and disease. Finally, it briefly introduces epigenetic regulation of phenotypes in Drosophila development and cancer through examples of homeotic gene regulation and implications for disease.

    Original Title

    10-19-05

    Copyright

    © Attribution Non-Commercial (BY-NC)

    Available Formats

    PPT, PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    This document discusses epigenetic regulation of phenotypes through genomic imprinting and DNA methylation. It provides an overview of genomic imprinting, describing the discovery of imprinting and key examples like Igf2/H19. It also discusses DNA methylation, covering topics like the basic concepts, techniques for studying methylation, functions of methylation, mechanisms of methylation-mediated repression, and relationships between methylation and disease. Finally, it briefly introduces epigenetic regulation of phenotypes in Drosophila development and cancer through examples of homeotic gene regulation and implications for disease.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    31 views53 pages

    Genomic Imprinting & Epigenetics: 徐国良 研究员 细胞大楼 300 电话 E-mail: [email protected]

    Uploaded by

    api-3700537
    This document discusses epigenetic regulation of phenotypes through genomic imprinting and DNA methylation. It provides an overview of genomic imprinting, describing the discovery of imprinting and key examples like Igf2/H19. It also discusses DNA methylation, covering topics like the basic concepts, techniques for studying methylation, functions of methylation, mechanisms of methylation-mediated repression, and relationships between methylation and disease. Finally, it briefly introduces epigenetic regulation of phenotypes in Drosophila development and cancer through examples of homeotic gene regulation and implications for disease.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    of 53

    Genomic Imprinting & Epigenetics

    基因印迹与表观遗传学

    徐国良 研究员
    细胞大楼 300
    电话 : 6433 5944
    E-mail: [email protected]
    Epigenetics October 17, 2005
    1. Introduction to Epigenetics
    1.1 Ubiquity of epigenetics
    1.2 Epigenetic Inheritance v.s. Mendelian inheritance
    1.3 Epigenetic gene regulation v.s. Signal transduction

    2. Genomic Imprinting
    2.1 Discovery
    2.2 Igf2/H19 paradigm
    2.3 Imprinting Center
    2.4 CTCF
    2.5 Evolutional aspects

    3. X-chromosome Inactivation
    Epigenetics October 19, 2005
    1. DNA methylation
    4.1 Basic concepts: methylation patterns, de novo methylation, maintenance methylation
    4.2 Techniques to study DNA methylation
    4.3 Biological functions of DNA methylation
    4.4 Mechanisms of DNA methylation-mediated transcriptional repression
    4.5 Methylation dynamics in development
    4.6 DNA methyltransferases
    4.7 DNA methylation and disease

    5. Epigenetic regulation of Phenotypes


    6.1 Epigenetic regulation of homeotic genes in the formation of Drosophila body plan
    6.2 Epigenetic regulation in the control of flowering
    6.3 Implications for cancer etc
    Mammalian DNA Methylation

    H2N H2N
    H CH3
    N N
    MTase
    SAM
    O N O N

    dC 5-methyl-dC
    CH3
    Methyltransferase in action
    Detection of genomic methylation by methylation-
    sensitive restriction enzymes

    HpaII, MspI MspI


    CH3

    -----CCGG----- -----CCGG-----
    -----GGCC----- -----GGCC-----
    CH3
    Detection of Methylcytosines: Bisulfite Sequencing
    DNA 甲基化的生物学功能

    • 遏制基因组内寄生虫（ intragenomic
    parasites ）
    • 控制基因活性状态 ( 开或关 )
    如在基
    因印迹 (genomic imprinting) 与 X 染色
    体失活 (X-inactivation) 过程中
    • 维持染色体结构稳定性
    • 维持正常胚胎发育

    • 甲基化与癌变和某些遗传病有关
    Review

    Science 293, 1068. August 10, 2001


    Bacterial Restriction/Modification Systems
    E. coli Dam methylation
    and Mismatch Repair

    CH3

    ---GATC---
    ---CTAG---
    CH3
    How DNA methylation works?

    Jones P. & Takai D. Science 293, 1068- (2001).


    DNA Methylation and Gene Silencing
    Methylation reprogramming
    in the germ line (A) and
    Preimplantation embryos (B)

    Reik W. et al, Science 293, p1089-


    Dynamic changes of DNA Methylation patterns in development
    Dnmt1 Protein and mRNA in Germ Cells and Early Embryos
    Mammalian DNA Methyltransferases
    Facial Features in ICF Syndrome
    A
    A­Miniou et al. Hum. Mol. Genet. 3(12):2093 (1994)
    B­Adapted from Brown et al. Hum. Genet. 96:411 (1995)
    DNMT3B mutations in ICF patients

    Ins744STP
    A766P
    158X G655S V810M
    S282P
    F/S 53 A585V L656T H814R

    I IV VI IX X
    ∆1aa
    6
    R54X Q204X V606A V718G D809G
    A603T V699G
    R832Q

    Xu et al., Nature, 1999;


    Hansen et al., PNAS, 1999;
    Shirohzu et al., Amer. J. Med. Genet., 2002.
    Conclusions

    • DNMT3B is the ICF disease gene


    • DNMT3B methylates satellite sequences
    • Cytosine methylation is essential for
    chromosome stability and heterochromatin
    formation.
    Diseases associated with abnormal
    methylation

    Diseases Genes involved


    ICF DNMT3B
    Rett MeCP2
    Fragile X Syndrome FMR
    α - Thalassaemia ATRX

    Cancer Tumor suppressor genes


    Azoospermia in Homozygous Males (Dnmt3L-/-)
    Abnormal development of embryos from homozygous females
    Loss of Maternal Methylation
    Imprints in Embryos Derived
    Homozygous Mother
    Biallelic Expression of Maternally Imprinted Genes in Dnmt3L -/+ Embryos
    Conclusions
    • Dnmt3L is essential for the establishment of
    maternal imprints (i.e. the de novo methylation
    of maternally imprinted genes in oogenesis)
    Cell 69, 915-926, 1992

    Targeted Mutation of the DNA Methyltransferase


    Results in Embryonic Lethality

    En Li, Timothy H. Bestor, and Rudolf Jaenisch


    Figure 1. Disruption of the 5’ End of the DNA MTase Locus
    Figure 2. Southern Blot Analysis of ES Cell Clones Targeted with pMT(N)neo or hyg
    Figure 3. Immunoblot Analysis of DNA MTase Figure 4. DNA MTase Enzyme Activity
    In Wild-Type and Mutant ES Cell Lines In Wild-Type and Mutant ES Cell Lines
    Figure 5. Quantitation of Genomic Figure 6. Southern Blot Analysis of DNA
    m5C Content Methylation of Endogenous Retroviral Geno
    In Wild-Type and Mutant ES Cell Lines
    In Wild-Type and Mutant ES Cell Lines
    wt

    Figure 7. Gross Morphology of Wild-Type


    and Mutant Embryos at 10.5 Days of
    Gestation
    mutant
    Figure 9. Immunoblot Analysis and m5C Quantitation of Wild-Type and Mutant Embryos

    ---C CGG---
    ---GGC C---
    Epigenetic Regulation of Phenotypes – Example 1

    Regulation of Drosophila Body Plans


    through Control of Homeotic Genes
    Polycomb, Trithorax, and Maintenance of Gene Expression
    A key feature of development in metameric animals is the definition of body
    segments where groups of cells with a specified fate will give rise to their relative
    body structures. Cell fates are specified by particular combinations of homeotic gene
    products. During early embryogenesis, maternal and segmentation genes regulate
    homeotic genes by binding to specific regulatory sequences located in the promoter
    regions.  Later in development, the expression pattern of homeotic genes as well as
    other important developmental genes are maintained by a cell memory system
    dependent on two groups of genes. The members of these two groups are able to
    recognize the active and inactive state of expression and fix it to the cell progeny
    through many cell divisions. These components have been classified in two genetic
    groups. The trithorax-group (trxG) maintain the active state of expression, while the
    Polycomb-group (PcG) counteracts this activation with a stable repressive function.
    There is strong evidence that the memory function encoded by these two groups
    of genes is achieved through regulation of higher order chromatin structures. PcG
    gene products form large multimeric protein complexes in Drosophila, mouse and
    human. PcG mediated gene silencing can be directed by DNA elements in cis, defined
    as PcG response elements (PRE). On the other hand, several trxG members act at
    elements defined as TRE (that overlap with PRE) via chromatin remodeling and
    induction of histone modifications that increase chromatin accessibility to
    基因表达状态的建立与维持需要
    transcription factors. :
    顺式调控元件 : specific DNA sequence
    反式作用因子 : DNA binding proteins including
    transcriptional factors
    Polycomb, Trithorax, and Maintenance of Gene Expression
    Early development  OFF ONMaternal, Gap, Pair­rule, Segment polarity
    Establishment of patterns Ubx

    OFF ON

    Polycomb­Group Maintenance phase trithorax­Group


    Transmission of 
    pattern after 
    disappearance of early 
    factors
    OFF ON

    Ubx OFF  Ubx ON

    wing  haltere 
    Update: December 2004
    PcG and trxG Proteins Associate to Multiple Genomic Loci
    DAPI Merge

    PH

    Polytene chromosome staining shows around 100 bands for each PcG protein
    PcG and trxG Proteins Bind to Specific DNA
    Elements, Named PRE and TRE

    PRE
    und by PcG proteins in vivo (in polytene chromosomes and by cross­linking experiments)

    inding leads to maintenance of PcG­dependent repression of reporter genes

    Repression is enhanced by homologous pairing of the transgenes

    TRE
    Bound by trxG proteins in vivo

    inding leads to maintenance of trxG­dependent activation of reporter genes

    PRE and TRE often overlap in the same genomic region
    Members of the PcG and of the trxG
    PcG Gene protein motif homologs
    Polycomb (Pc) chromo domain M33 (mouse); hPC (human)
    (Binding to H3 methyl K9 or K27)
    PRC1  polyhomeotic (ph) one zinc finger Mph1/Rae­28 (mouse); hph1; hph2 (human)
    comple
    x Posterior sex combs (Psc) RING finger bmi­1 (mouse/human); mel­18 (mouse)

    Esc/E(z) Enhancer of zeste (E(z)) SET (H3MTase) Ezh1; Ezh2 (human); clf (Arabidopsis)


    Complex extra sex combs (esc) WD repeat Eed (mouse); hEED (human)

    Pleiohomeotic (pho) Zinc­finger (DNA binding) hYY­1 (human); mYY­1 (mouse)

    trxG Gene protein motif homologs


    TAC1  trithorax (trx) SET  (H3 HMTase)/ PHD­finger MLL/ALL­1/HRX (human)
    complex
    Ash­1 SET  (H3/H4HMTase)/ PHD­finger ASH­1 (human); NSD1 (mouse)

    bromo domain SWI2/SNF2 (yeast)
    Brm complex brahma (brm)
    (DNA dependent ATPase/helicase) brg1 (mouse/human); Hbrm (human)

    FACT  Trithorax­like (Trl) BTB/POZ (dimerization)


    complex zinc finger (DNA binding)
    Action of PcG and trxG Complexes on Chromatin
    Nucleosome trxG
    remodeling
    Ac (BRM complex) ON Maintenance of
    Histone active states
    acetylation and (open
    methylation
    (TAC1 and Me K4 H3 chromatin)
    ASH1
    complexes)

    Target
    PRE gene

    OFF PcG
    Deacetylation and Me K27 H3
    methylation Maintenance of
    (ESC-E(Z) repressed states
    complex) (compact
    - Chromatin chromatin)
    compaction Ub H2A
    - H2A
    Ubiquitination
    (PRC1 complex)
    Histone H3 methylation and Polycomb
    Pc H3 K9  Merge
    triMe

    Data from: 
    Ringrose et al. 
    (2004) Mol. Cell 
    16, 641 Pc H3 K27  Merge
    triMe
    There is a strong but not absolute correlation between trimethylation of K27 (and
    K9) trimethylation and Polycomb recruitment at target loci. i.e. there is more to Pc
    recruitment
    Epigenetic Regulation of Phenotypes – Example 2

    Regulation in the Control of Flowering


    Vernalization Requirement and Response in Crops and Arabidopsis
    “ 春化作用”后才能开花
    The Role of FLC in Vernalization Requirement and Response
    VRN Genes Are Required to Maintain Suppression of FLC after Vernalization

    fca-1

    vrn1fca-1
    Epigenetic Silencing of FLC

    You might also like