Imprinted Segments in the Human Genome: Different Dna Methylation Patterns in the Prader-Willi/Angelman Syndrome Region As Determined by the Genomic Sequencing Method

Zeschnigk, Michael; Schmitz, Birgit; Dittrich, Bärbel; Buiting, Karin; Horsthemke, Bernhard; Doerfler, Walter

doi:10.1093/hmg/6.3.387

Cited by 130 publications

(89 citation statements)

References 37 publications

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“…In the germline, methyl groups are attached to or removed from the CGrich imprinting region in a paternal-or maternal-specific pattern. The CG-rich imprinting region in chromosome 15q11-q13 is a 300 bp segment (Zeschnigk et al, 1997) that is called the Prader-Willi syndrome imprinting center (PWS-IC) (Fig. 2).…”

Section: Genomic Imprinting and Angelman Syndromementioning

confidence: 99%

“…The PWS-IC is located in the upstream promoter region of the SNURF/SNRPN transcript (Sutcliffe et al, 1994;Zeschnigk et al, 1997;Bielinska et al, 2000;ElMaarri et al, 2001). The PWS-IC is one part of a bipartite imprinting regulatory element; the second component is located 35-40 kb upstream of PWS-IC and is referred to as the AS imprinting center (AS-IC) (Dittrich et al, 1996).…”

Section: Genomic Imprinting and Angelman Syndromementioning

confidence: 99%

See 1 more Smart Citation

Angelman Syndrome, a Genomic Imprinting Disorder of the Brain

Chamberlain¹,

Lalande²

2010

J. Neurosci.

101

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Section: Genomic Imprinting and Angelman Syndromementioning

confidence: 99%

Section: Genomic Imprinting and Angelman Syndromementioning

confidence: 99%

Angelman Syndrome, a Genomic Imprinting Disorder of the Brain

Chamberlain¹,

Lalande²

2010

J. Neurosci.

101

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“…6,14 Quantitative analysis of methylated alleles Bisulfite treatment: the procedure was modified from established protocols. 18 Genomic DNA (1 mg in 30 ml) was denatured by adding 3 ml freshly prepared 3 M NaOH and incubating the solution at 371C for 15 min. For complete denaturation, the samples were incubated at 951C for 1 min and immediately cooled on ice.…”

Section: Methylation Analysismentioning

confidence: 99%

IGF2/H19 hypomethylation in Silver–Russell syndrome and isolated hemihypoplasia

et al. 2008

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Silver-Russell syndrome (SRS) is a clinically and genetically heterogeneous syndrome characterized by severe pre and postnatal growth retardation, body asymmetry and a typical facial phenotype with a triangular face and relative macrocephaly. In 30% of patients, the differentially methylated IGF2/H19 imprinting center region (ICR1) on chromosome 11p15 was found to be hypomethylated, as determined by Southern blot analysis of an HpaII restriction site close to the third CTCF-binding site (CTS3) within ICR1. Using bisulfite treatment and a real-time PCR-based methylation assay (QAMA), we analyzed the third and sixth CTCF-binding sites (CTS3, CTS6) in 5 patients with CTS3 hypomethylation, in 14 patients who were suspected to have SRS but were normal by Southern blot analysis, and in 1 patient with body asymmetry without any other features of SRS or Beckwith-Wiedemann syndrome (BWS). In all 5 patients with CTS3 hypomethylation, in 5 of 14 patients who were judged to be normal at CTS3 by Southern blot analysis and in the patient with isolated body asymmetry, we found CTS3 and CTS6 hypomethylation by QAMA. Using methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA), we obtained similar results at four additional ICR1 sites in the CTS6 region. These results show that ICR1 hypomethylation occurs more often in SRS patients than as previously thought as well as in isolated hemihypoplasia. Furthermore, we show that methylation analysis by QAMA and MLPA is more sensitive in detecting ICR1 hypomethylation than Southern blot analysis of CTS3.

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“…Genomic DNA from PHA-stimulated human lymphocytes, human skin fibroblasts, and gorilla lymphoblastoid cells was purified and treated with sodium bisulfite according to standard methods. Methylation-specific PCR analysis of the SNURF-SNRPN exon 1/promoter region was performed with primers described by Zeschnigk et al (47). -385H1, RP11-125E1, RP11-512F9, RP11-131I21, RP11-145P16, RP11-183K8, RP11-466L14, RP11-259N18, RP11-2C7, RP11-446P9, RP11-71L8, RP11-272I21, RP11-75O6, RP11-345N11, RP11-142M24, RP11-147D1, RP11-188P24, RP11-570N16, RP11-321B18, RP11-100M12, RP11-107D24, RP11-30G8) were ordered at BACPAC Resources Center (Oakland, CA); DNA preparation was performed by using alkaline lysis according to standard protocols.…”

Section: Methodsmentioning

confidence: 99%

Maintenance of imprinting and nuclear architecture in cycling cells

Teller

Solovei

Buiting

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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(14) provided a first case-in-point. They performed FISH experiments in normal, phytohemagglutinin (PHA)-stimulated human lymphocytes with a probe covering a segment of the Angelman syndrome/Prader-Willi syndrome (AS/PWS) locus on HSA 15q11-13 and measured 3D interdistances between the two signals in samples of 50 nuclei at different interphase stages. Approximately 38% of nuclei at G 1 , Ϸ18% at early S phase, Ϸ58% at late S phase, and Ϸ18% at G 2 revealed distances Յ2 m, which were considered as a proximity criterion for a possible functional interaction between the two AS/PWS loci. Notably, the significant increase of associations at late S phase was not found in cells from PWS and AS patients. Furthermore, LaSalle and Lalande (14) mentioned that they obtained similar results for the Beckwith-Wiedemann syndrome (BWS) locus, a second imprinted locus on HSA 11p15.5. Based on these findings, LaSalle and Lalande formulated the kissing hypothesis, which states that a transient spatial association between the oppositely imprinted regions during late S phase is a necessary condition to maintain the imprinted status of cycling cells from one cell cycle to the next. This hypothesis was widely cited and further supported by a 2D FISH study from Riesselmann and Haaf (15), who measured 2D interhomolog distances between oppositely imprinted loci on MMU 7 as a fraction of the nuclear diameter in flattened mouse fibroblast nuclei obtained in cytospin preparations. Somatic pairing was assumed when hybridization signals apparently overlapped or touched each other. By this criterion, significantly higher values were found for BrdU-positive nuclei (13%; 27/208 nuclei) compared with BrdUnegative nuclei (9.2%; 21 of 228 nuclei). No increase of pairing frequencies during S phase was found for control regions not subject to imprinting. In contrast, Nogami et al. (16), who studied the frequency of associations between the two AS loci with probes for the imprinted SNRPN alleles in 3D preserved interphase nuclei of human myeloid leukemia HL60 cells, did not find evidence to further support the hypothesis.Importantly, a large variability of 3D positions was noted for pairs of HSA 11 and HSA 15 chromosome territories (CTs) both within nuclei and between nuclei of different cells. Given this variability, a transient kissing event during late S phase between loci widely separated at other stages of interphase requires specific, long-range, directed chromatin movements. The present study was initiated with the intent to clarify whether such movements lead to the transient juxtaposition of entire homologous CTs or, alternatively, whether homologous CTs stay in place, whereas giant loops harboring genes poised for a kissing event expand from territory surfaces and meet each other in between (1,6,8,17). We visualized the two AS/PWS regions in PHA-stimulated lymphocytes by 3D-FISH with a BAC contig covering most of the AS/PWS locus and measured the 3D distances between the two regions in G 1 , early, mid, and late S phase, G 2 , as well as in...

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Imprinted Segments in the Human Genome: Different Dna Methylation Patterns in the Prader-Willi/Angelman Syndrome Region As Determined by the Genomic Sequencing Method

Cited by 130 publications

References 37 publications

Angelman Syndrome, a Genomic Imprinting Disorder of the Brain

Angelman Syndrome, a Genomic Imprinting Disorder of the Brain

IGF2/H19 hypomethylation in Silver–Russell syndrome and isolated hemihypoplasia

Maintenance of imprinting and nuclear architecture in cycling cells

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