Assessment of a rapid-cycle PCR assay for the identification of the recurrent c.3421C&gt;T mutation in the ABCC6 gene in pseudoxanthoma elasticum patients

Götting, Christian; Schulz, Veronika; Hendig, Doris; Grundt, Alexander; Dreier, Jens; Szliska, Christiane; Brinkmann, Thomas; Kleesiek, Knut

doi:10.1038/sj.labinvest.3700004

Cited by 10 publications

(6 citation statements)

References 25 publications

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“…Among the 22 ABCC6 sequence variations found in this study, 17 had been previously described as PXE-causing mutations and ABCC6 polymorphisms [16, 19,22,23,24, 30, 35, 36], and 5 were novel DNA alterations resulting in the missense and silent variants p.Q655R, p.P664S, p.R1114C, p.G1311E and p.R1418R. An alignment using the ClustalW program showed that the missense mutations alter amino acid residues conserved in the human, mouse and rat MRP6 proteins.…”

Section: Resultsmentioning

confidence: 99%

“…Among these, 3 variants (p.R724K, p.V725V and p.I742V) had already been described as ABCC6 polymorphisms [36]. 14 DNA alterations had been shown to be PXE-causing mutations [16, 19,22,23,24, 30, 35, 36] and, consequently, have not been further examined in this study. Two ABCC6 sequence alterations were novel, namely the alterations p.R1114C and p.G1311E.…”

Section: Discussionmentioning

confidence: 99%

“…The restriction enzyme Esp3 I (MBI Fermentas, St. Leon-Rot, Germany) was used for the detection of the mutations c.3340C>T, c.3341G>A and c.3389C>T and Alu I (Gibco BRL, Gaithersburg, Md., USA) for the allele-specific genotyping of the mutation c.3932G>A. Digested and undigested products were subjected to agarose gel electrophoresis and visualized after ethidium bromide staining. The c.3421C>T, c.2175A>T, c.2224A>G, c.4209C>A and Ex23_Ex29del PCR and RFLP genotyping was performed as described previously [35,36,37]. DHPLC analysis of the PCR products E16, E17, E18, E29a and E30 was used to demonstrate the presence or absence of DNA variations in healthy controls.…”

Section: Methodsmentioning

confidence: 99%

“…To amplify the exons 1–31 of the ABCC6 gene, ABCC6 -specific primers were designed in the flanking introns to amplify DNA fragments encompassing the exon/intron boundaries and the exon. The PCR reactions were performed as described previously [35] with fragment-specific annealing temperatures. The primer sequences, the annealing temperatures and the sizes of the PCR products of the DHPLC-analyzed ABCC6 exons (16–18, 24 and 28–30) are given in table 2.…”

Section: Methodsmentioning

confidence: 99%

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Analysis of Sequence Variations in the <i>ABCC6</i> Gene among Patients with Abdominal Aortic Aneurysm and Pseudoxanthoma Elasticum

Schulz

Hendig

Schillinger³

et al. 2005

J Vasc Res

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Abdominal aortic aneurysm (AAA) is characterized by dilatation of arterial walls, which is accompanied by degradation of elastin and collagen molecules. Biochemical and environmental factors are known to be relevant for AAA development, and familial predisposition is well recognized. A connective tissue disorder that is also associated with fragmentation of elastic fibers is Pseudoxanthoma elasticum (PXE). PXE is caused by mutations in the ABCC6 gene and mainly affects dermal, ocular and all vascular tissues. To investigate whether variations in ABCC6 are found in AAA patients and to determine mutations in PXE patients, we analyzed seven selected ABCC6 exons of 133 AAA and 54 PXE patients subjected to mutational analysis. In our cohort of AAA patients, we found five ABCC6 alterations, which result in missense or silent amino acid variants. The allelic frequencies of these sequence variations were not significantly different between AAA patients and healthy controls. Therefore, we suggest that alterations in ABCC6 are not a genetic risk factor for AAA. Mutational screening of the PXE patients revealed 19 different ABCC6 variations, including two novel PXE-causing mutations. These results expand the ABCC6 mutation database in PXE.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Analysis of Sequence Variations in the <i>ABCC6</i> Gene among Patients with Abdominal Aortic Aneurysm and Pseudoxanthoma Elasticum

Schulz

Hendig

Schillinger³

et al. 2005

J Vasc Res

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“…[2][3][4] To date, more than 120 PXE causing mutations have been identified. [3][4][5][6][7][8][9] The central hallmark of PXE is degenerative calcification with subsequent disintegration and destruction of elastic tissue in the skin, eyes, and cardiovascular system. PXE shows considerable clinical heterogeneity.…”

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confidence: 99%

Polymorphisms in the xylosyltransferase genes cause higher serum XT-I activity in patients with pseudoxanthoma elasticum (PXE) and are involved in a severe disease course

Schön

Schulz²,

Prante³

et al. 2006

Journal of Medical Genetics

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Background: Pseudoxanthoma elasticum (PXE) is a heritable connective tissue disorder caused by mutations in the ABCC6 gene. Fragmentation of elastic fibres and deposition of proteoglycans result in a highly variable clinical picture. The altered proteoglycan metabolism suggests that enzymes from this pathway function as genetic co-factors in the severity of PXE. Therefore, we propose the XYLT genes encoding xylosyltransferase I (XT-I) as the chain-initiating enzyme in the biosynthesis of proteoglycans and the highly homologous XT-II as potential candidate genes. Methods: We screened all XYLT exons in 65 German PXE patients using denaturing high performance liquid chromatography and analysed the influence of the variations on clinical characteristics. Results: We identified 22 variations in the XYLT genes. The missense variation p.A115S (XT-I) is associated with higher serum XT activity (p = 0.005). The amino acid substitution p.T801R (XT-II; c.2402C.G) occurs with significantly higher frequency in patients under 30 years of age at diagnosis (43% v 26%; p = 0.04); all PXE patients with this variation suffer from skin lesions compared to only 75% of the wild type patients (p = 0.002). c.166G.A, c.1569C.T, and c.2402C.G in the XYLT-II gene were found to be more frequent in patients with higher organ involvement (p = 0.04, p = 0.01, and p = 0.02, respectively). Conclusions: Here we show for the first time that variations in the XYLT-II gene are genetic co-factors in the severity of PXE. Furthermore, the higher XT activity in patients with the exchange p.A115S (XT-I) indicates that this polymorphism is a potential marker for increased remodelling of the extracellular matrix.

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New ABCC6 gene mutations in German pseudoxanthoma elasticum patients

et al. 2004

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Pseudoxanthoma elasticum (PXE; OMIM 177850 and 264800) is a rare heritable disorder of the connective tissue affecting the extracellular matrix of the skin, eyes, gastrointestinal system, and cardiovascular system. It has recently been found that mutations in the ABCC6 gene encoding the multidrug resistance-associated protein (MRP) 6 cause PXE. This study examined novel mutations in the ABCC6 gene in our cohort of 76 German PXE patients and 54 unaffected or not yet affected relatives with a view to expanding the known mutational spectrum of the gene. Mutational analysis was performed using denaturing high-performance liquid chromatography and direct sequencing. The mutational screening revealed a total of 22 different ABCC6 sequence variations. We identified seven novel and four previously described PXE-associated mutations as well as eight novel neutral ABCC6 sequence variants. The new PXE-associated mutations included five missense mutations, one single base pair deletion, and one larger out-of-frame deletion. We suspect that the novel missense mutations lead to an impaired function of MRP6. Both deletions are predicted to result in a dysfunctional MRP6 protein. The seven new ABCC6 mutations were not present in 200 alleles from healthy blood donors which served as a control cohort. Most of the PXE patients who were found to carry PXE-causing ABCC6 mutations were assumed to manifest the PXE phenotype because of a compound heterozygous genotype. However, a genotype-phenotype correlation could not be established for the detected ABCC6 mutations. In summary, our data give a further insight into the spectrum of ABCC6 mutations in PXE patients.

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Assessment of a rapid-cycle PCR assay for the identification of the recurrent c.3421C>T mutation in the ABCC6 gene in pseudoxanthoma elasticum patients

Cited by 10 publications

References 25 publications

Analysis of Sequence Variations in the <i>ABCC6</i> Gene among Patients with Abdominal Aortic Aneurysm and Pseudoxanthoma Elasticum

Analysis of Sequence Variations in the <i>ABCC6</i> Gene among Patients with Abdominal Aortic Aneurysm and Pseudoxanthoma Elasticum

Polymorphisms in the xylosyltransferase genes cause higher serum XT-I activity in patients with pseudoxanthoma elasticum (PXE) and are involved in a severe disease course

New ABCC6 gene mutations in German pseudoxanthoma elasticum patients

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