AnEcoRV polymorphism in exon 40 of the tuberous sclerosis 2 (TSC2) gene

Bakel, I. Van; Sepp, Tiina; Green, Andrew J.

doi:10.1006/mcpr.1996.0080

Cited by 6 publications

(3 citation statements)

References 2 publications

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“…At the TSC1 locus the markers tested were: exon 14 (1556 A/G); intron 21 (2847-4 {A} 17-21 ), exon 22 (3050 C/T) ) and PM1, PM2, PM4 and PM5 (this study). The TSC2 locus markers were Kg8 (Snarey et al 1994); exon 40 (1734 T/C) (van Bakel et al 1997); IVS8 29 bp VNTR (Migone 1998), EJ1 (Qian et al 1996) and LP1, LP7 and LP10 (this study). PCR amplification of tumour and constitutional DNA samples was carried out in parallel in 96 well microtitre plates (Hybaid).…”

Section: Methodsmentioning

confidence: 98%

Molecular analysis of the TSC1 and TSC2 tumour suppressor genes in sporadic glial and glioneuronal tumours

et al. 2000

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Reduced expression of the TSC2 tumour suppressor gene product, tuberin, has been reported in sporadic astrocytomas, suggesting that the TSC genes may play a role in formation of sporadic glial or glioneuronal tumours. We studied paired constitutional and tumour DNA samples from 100 patients with sporadic glial and glioneuronal tumours for loss of heterozygosity (LOH) at the TSC1 and TSC2 loci using a combination of seven previously reported and seven novel polymorphic markers. LOH was seen in 1/16 astrocytomas, 3/15 ependymomas, 5/16 gangliogliomas, 2/14 glioblastoma multiforme, 0/7 oligodendrogliomas, 0/7 tumours of mixed oligodendrocytic/astrocytic histology, 2/11 pilocytic astrocytomas and 0/1 subependymal giant cell astrocytomas informative at both loci. However, SSCP screening of all coding exons of the TSC1 or TSC2 genes in the tumours displaying LOH, and of both genes in 21 gangliogliomas, revealed no intragenic mutations. The lack of demonstrable inactivation of both alleles of either TSC gene in any of the tumours investigated suggests that they do not play a frequent role in the aetiology of sporadic glial or glioneuronal tumours.

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Section: Methodsmentioning

confidence: 98%

Molecular analysis of the TSC1 and TSC2 tumour suppressor genes in sporadic glial and glioneuronal tumours

et al. 2000

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“…The pathogenicity of several missense changes identified in that study remains unconfirmed. van Bakel et al (1997) have used the protein-truncation test to study 18 unrelated patients and have identified truncating mutations in 5 of them. Au et al (1998) have used SSCP analysis of all coding exons to study 90 unrelated cases and have identified 22 putative mutations.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive Mutation Analysis of TSC1 and TSC2—and Phenotypic Correlations in 150 Families with Tuberous Sclerosis

Jones

Shyamsundar

Thomas

et al. 1999

The American Journal of Human Genetics

464

353

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Tuberous sclerosis (TSC [MIM 191090 and MIM 191100]) is an autosomal dominant disorder characterized by hamartomas in many organs. Two thirds of cases are sporadic and are thought to represent new mutations. TSC is caused by mutations affecting either of the presumed tumor-suppressor genes, TSC1 and TSC2. Both appear to function as tumor suppressors, because somatic loss or intragenic mutation of the corresponding wild-type allele is seen in the associated hamartomas. Here we report the first comprehensive mutation analysis of TSC1 and TSC2 in a cohort of 150 unrelated TSC patients and their families, using heteroduplex and SSCP analysis of all coding exons and using pulsed-field gel electrophoresis and conventional Southern blot analysis and long PCR to screen for large rearrangements. Mutations were characterized in 120 (80%) of the 150 cases, affecting TSC1 in 22 cases and TSC2 in 98 cases. TSC1 mutations were significantly underrepresented in sporadic cases (P=. 000185). Twenty-two patients had TSC2 missense mutations that were found predominantly in the GAP-related domain (eight cases) and in a small region encoded in exons 16 and 17, between nucleotides 1849 and 1859 (eight cases), consistent with the presence of residues performing key functions at these sites. In contrast, all TSC1 mutations were predicted to be truncating, consistent with a structural or adapter role for the encoded protein. Intellectual disability was significantly more frequent in TSC2 sporadic cases than in TSC1 sporadic cases (P=.0145). These data provide the first representative picture of the distribution and spectrum of mutations across the TSC1 and TSC2 loci in clinically ascertained TSC and support a difference in severity of TSC1- and TSC2-associated disease.

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“…Genomic DNA isolated from whole blood and unsorted or sorted cells was amplified by PCR for the exon 40 polymorphism (T5202C; rs1748) (28,29) or for a splice site polymorphism (C482-3T; rs1800720) (28,30), using these primer sequences: TSC40S-Hex, 59-Hex-ATGGAG GGCCTTGTGGACAC-39, and TSC40AS, 59-CGGAGCCGCTTGA TGTG-39; TSCspliceS, 59-GGAGATGTAGATTCGGCGTC-39, and TSCspliceAS-Hex, 59-Hex-CTGCGGAGCTGAACTTAGG-39. PCR products were digested with the appropriate restriction enzyme (EcoRV for T5151C; PvuII for C482-3T) (New England Biolabs, Beverly, MA), and then analyzed with a 3100 Genetic Analyzer (Applied Biosystems).…”

Section: Single-nucleotide Polymorphism Analysismentioning

confidence: 99%

Phenotypic Characterization of Disseminated Cells with TSC2 Loss of Heterozygosity in Patients with Lymphangioleiomyomatosis

Cai¹,

Pacheco–Rodriguez²,

Fan³

et al. 2010

Am J Respir Crit Care Med

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Rationale: Lymphangioleiomyomatosis (LAM), occurring sporadically (S-LAM) or in patients with tuberous sclerosis complex (TSC), results from abnormal proliferation of LAM cells exhibiting mutations or loss of heterozygosity (LOH) of the TSC genes, TSC1 or TSC2. Objectives: To identify molecular markers useful for isolating LAM cells from body fluids and determine the frequency of TSC1 or TSC2 LOH. Methods: Candidate cell surface markers were identified using gene microarray analysis of human TSC2 2/2 cells. Cells from bronchoalveolar lavage fluid (BALF), urine, chylous effusions, and blood were sorted based on reactivity with antibodies against these proteins (e.g., CD9, CD44v6) and analyzed for LOH using TSC1-and TSC2-related microsatellite markers and single nucleotide polymorphisms in the TSC2 gene. Measurements and Main Results: CD44v6 1 CD9 1 cells from BALF, urine, and chyle showed TSC2 LOH in 80%, 69%, and 50% of patient samples, respectively. LAM cells with TSC2 LOH were detected in more than 90% of blood samples. LAM cells from different body fluids of the same patients showed, in most cases, identical LOH patterns, that is, loss of alleles at the same microsatellite loci. In a few patients with S-LAM, LAM cells from different body fluids differed in LOH patterns. No patients with S-LAM with TSC1 LOH were identified, suggesting that TSC2 abnormalities are responsible for the vast majority of S-LAM cases and that TSC1-disease may be subclinical. Conclusions: Our data support a common genetic origin of LAM cells in most patients with S-LAM, consistent with a metastatic model. In some cases, however, there was evidence for genetic heterogeneity between LAM cells in different sites or within a site.

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AnEcoRV polymorphism in exon 40 of the tuberous sclerosis 2 (TSC2) gene

Cited by 6 publications

References 2 publications

Molecular analysis of the TSC1 and TSC2 tumour suppressor genes in sporadic glial and glioneuronal tumours

Molecular analysis of the TSC1 and TSC2 tumour suppressor genes in sporadic glial and glioneuronal tumours

Comprehensive Mutation Analysis of TSC1 and TSC2—and Phenotypic Correlations in 150 Families with Tuberous Sclerosis

Phenotypic Characterization of Disseminated Cells with TSC2 Loss of Heterozygosity in Patients with Lymphangioleiomyomatosis

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