Identification and Detection of the Common 65-kb Deletion Breakpoint in the Nephropathic Cystinosis Gene (CTNS)

Anikster, Yair; Lucero, Cynthia; Touchman, Jeffrey W.; Huizing, Marjan; McDowell, Geraldine; Shotelersuk, Vorasuk; Green, Eric D.; Gahl, William A.

doi:10.1006/mgme.1998.2790

Cited by 78 publications

(76 citation statements)

References 25 publications

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“…This method has been employed to detect hemizygosity for other disorders (35,36). We chose the CTNS and the HPS3 genes as internal control genes for co-amplification because we had DNA from patients hemizygous for these genes (24,25,37). We co-amplified HPS5 exon 17 or exon 22 with CTNS exon 3 ( Figures 4A,B) and HPS5 exon 17 with HPS3 upstream of exon 1 ( Figure 4C).…”

Section: Gene-dosage Multiplex Pcrmentioning

confidence: 99%

“…CTNS (cystinosin) (GI: 4826681) exon 3 (200-bp) was amplified using forward primer 5 0 -CAGGGAGCT-GAGCTGATTCA-3 0 and reverse primer 5 0 -CTCCTTCC-TCTAGCCACCAT-3 0 , and an HPS3 (GI: 15088620) 397-bp gDNA fragment upstream of exon 1 was amplified using forward primer 5 0 -CGTGAACTCCACGTTGAGATGTCA-3 0 , and reverse primer 5 0 -CGTTCTGACAATTCATCATCTATC-3 0 . Genomic DNA from a normal control, patient #46, patient #48, a cystinosis patient hemizygous for the common 57.3-kb CTNS deletion (including exon 3) (37), and an HPS-3 patient hemizygous for the common 3.9-kb central Puerto-Rican HPS3 deletion (patient #90 in (25)) were used as templates for gene-dosage multiplex PCR. Three PCR experiments with a combination of two primer sets were performed: HPS5 exon 17 combined with CTNS exon 3; HPS5 exon 22 combined with CTNS exon 3; and HPS5 exon 17 combined with HPS3 upstream of exon 1.…”

Section: Molecular Analysismentioning

confidence: 99%

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Cellular, Molecular and Clinical Characterization of Patients with Hermansky–Pudlak Syndrome Type 5

Huizing

Hess

Dorward

et al. 2004

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Hermansky-Pudlak syndrome (HPS) is a disorder of lysosome-related organelles such as melanosomes and platelet dense granules. Seven genes are now associated with HPS in humans. An accurate diagnosis of each HPS subtype has important prognostic and treatment implications. Here we describe the cellular, molecular, and clinical aspects of the recently identified HPS-5 subtype. We first analyzed the genomic organization and the RNA expression pattern of HPS5, located on chromosome 11p14, and demonstrated tissue-specific expression of at least three alternatively spliced HPS5 mRNA transcripts, coding for HPS5A and HPS5B proteins, that differ at their 5 0 -ends. Genetic screening of 15 unassigned HPS patients yielded six new HPS5 mutations in four patients. Clinically, our HPS-5 patients exhibited iris transillumination, variable hair and skin pigmentation, and absent platelet dense bodies, but not pulmonary fibrosis or granulomatous colitis. In two patients with homozygous missense mutations, hemizygosity was ruled out by gene-dosage multiplex polymerase chain reaction, and immunocytochemical analyses of their fibroblasts supported the HPS-5 diagnosis. Specifically, LAMP-3 distribution was restricted to the perinuclear region in HPS-5 fibroblasts, in contrast to the normal LAMP-3 distribution, which extended to the periphery. This specific intracellular vesicle distribution in fibroblasts, in combination with the clinical features, will improve the characterization of the HPS-5 subtype.

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Section: Gene-dosage Multiplex Pcrmentioning

confidence: 99%

Section: Molecular Analysismentioning

confidence: 99%

Cellular, Molecular and Clinical Characterization of Patients with Hermansky–Pudlak Syndrome Type 5

Huizing

Hess

Dorward

et al. 2004

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“…The most common mutation causing the infantile nephropathic form of cystinosis is a 57-kb deletion involving most of the CTNS gene (Touchman et al, 2000;Town et al, 1998). Patients with this deletion have an identical 5'-breakpoint upstream and a 3´-breakpoint in exon 10 of the gene (Anikster et al, 1999a;Forestier et al, 1999). In addition to this most frequent alteration, smaller deletions, missense mutations, and frameshifts have been described in European and Americanbased populations of cystinosis patients leading to downstream stop codons or abolition of splice sites.…”

Section: Introductionmentioning

confidence: 99%

Analysis of the CTNS gene in patients of German and Swiss origin with nephropathic cystinosis

et al. 2002

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The autosomal recessive lysosomal storage disorder, nephropathic cystinosis is characterized by impaired transport of free cystine out of lysosomes. The gene responsible for cystinosis, CTNS, consists of 12 exons and encodes a 55 kDa putative lysosomal membrane protein, called cystinosin. Up to now more than 55 different CTNS mutations have been described in cystinosis. We have analyzed the mutation pattern in a population of 40 cystinosis patients from 35 families of German and Swiss origin. CTNS mutations in 68 out of 70 alleles were identified. The common 57-kb deletion accounted for 65% of the alleles. In five patients we found a known GACT deletion at position 18-21. In two patients we identified a nucleotide substitution at codon 339 and one patient showed a CG insertion at position 697-698. In five patients we observed a G insertion at position 926-927. Moreover, five novel mutations including two deletions involving exon 3 (61-61+2delGGT) and exon 6 (280delG), two insertions in exon 6 (292-293insA) and exon 7 (684insCACTT) and one nucleotide substitution in exon 11 (923G>T) have been identified. These data provide a basis for routine molecular diagnosis of cystinosis in the central European population, especially in cystinosis patients of German and Swiss origin.

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“…A number of cystinosis-causing CTNS mutations have now been reported (Shotelersuk et al 1998a;Town et al 1998). The most prevalent mutation reported to date is a large (>55-kb) deletion, with 33%-44% of affected patients being homozygous for this deletion (Town et al 1998;Anikster et al 1999). In addition, at least 11 other smaller disease-causing deletions have been reported (Shotelersuk et al 1998a;Forestier et al 1999), suggesting that this genomic region may be prone to rearrangement.…”

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The Genomic Region Encompassing the Nephropathic Cystinosis Gene (CTNS): Complete Sequencing of a 200-kb Segment and Discovery of a Novel Gene within the Common Cystinosis-Causing Deletion

et al. 2000

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Nephropathic cystinosis is an autosomal recessive disorder caused by the defective transport of cystine out of lysosomes. Recently, the causative gene (CTNS) was identified and presumed to encode an integral membrane protein called cystinosin. Many of the disease-associated mutations in CTNS are deletions, including one >55 kb in size that represents the most common cystinosis allele encountered to date. In an effort to determine the precise genomic organization of CTNS and to gain sequence-based insight about the DNA within and flanking cystinosis-associated deletions, we mapped and sequenced the region of human chromosome 17p13 encompassing CTNS. Specifically, a bacterial artificial chromosome (BAC)-based physical map spanning CTNS was constructed by sequence-tagged site (STS)-content mapping. The resulting BAC contig provided the relative order of 43 STSs. Two overlapping BACs, which together contain all of the CTNS exons as well as extensive amounts of flanking DNA, were selected and subjected to shotgun sequencing. A total of 200,237 bp of contiguous, high-accuracy sequence was generated. Analysis of the resulting data revealed a number of interesting features about this genomic region, including the long-range organization of CTNS, insight about the breakpoints and intervening DNA associated with the common cystinosis-causing deletion, and structural information about five genes neighboring CTNS (human ortholog of rat vanilloid receptor subtype 1 gene, CARKL, P2X5, and HUMINAE). In particular, sequence analysis detected the presence of a novel gene (CARKL) residing within the most common cystinosis-causing deletion. This gene encodes a previously unknown protein that is predicted to function as a carbohydrate kinase. Interestingly, both CTNS and CARKL are absent in nearly half of all cystinosis patients (i.e., those homozygous for the common deletion).

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Identification and Detection of the Common 65-kb Deletion Breakpoint in the Nephropathic Cystinosis Gene (CTNS)

Cited by 78 publications

References 25 publications

Cellular, Molecular and Clinical Characterization of Patients with Hermansky–Pudlak Syndrome Type 5

Cellular, Molecular and Clinical Characterization of Patients with Hermansky–Pudlak Syndrome Type 5

Analysis of the CTNS gene in patients of German and Swiss origin with nephropathic cystinosis

The Genomic Region Encompassing the Nephropathic Cystinosis Gene (CTNS): Complete Sequencing of a 200-kb Segment and Discovery of a Novel Gene within the Common Cystinosis-Causing Deletion

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