PAH Gene Mutations in the Sicilian Population: Association with Minihaplotypes and Expression Analysis

Mirisola, Mario G.; Calı̀, Francesco; Gloria, Angelo; Schinocca, Pietro; D'Amato, Monica; Cassarà, Giuseppina; Leo, Giacomo De; Palillo, Letizia; Meli, Concetta; Romano, Valentino

doi:10.1006/mgme.2001.3211

Cited by 15 publications

(6 citation statements)

References 28 publications

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“…DNA samples for the patients were already available in the DNA bank of hyperphenylalaninemia patients located at the OASI Institute (Troina). The 43 patients were selected for PAH gene exon deletion screening because either one or both PAH mutant alleles remained unknown following previous DGGE/DHPLC and DNA sequencing analyses performed on a total of 802 alleles (Mirisola et al, 2001), [F. Calì and C. Carducci unpublished data].…”

Section: Methods Patientsmentioning

confidence: 99%

“…pahdb.mcgill.ca). Nevertheless, in many of these studies, the mutant genotype has remained partially or completely undetermined in about less than 10% of patients (Mirisola et al, 2001). In recent years, several studies (Gable et al, 2003;Desviat et al, 2006;Kozak et al, 2006;Birk Moller et al, 2007;Lee et al, 2008) have shown that a significant proportion of these undetermined alleles consist of large deletions overlapping one or more exons.…”

Section: Introductionmentioning

confidence: 99%

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Exon deletions of the phenylalanine hydroxylase gene in Italian hyperphenylalaninemics

et al. 2010

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A consistent finding of many studies describing the spectrum of mutant phenylalanine hydroxylase (PAH) alleles underlying hyperphenylalaninemia is the impossibility of achieving a 100% mutation ascertainment rate using conventional gene-scanning methods. These methods include denaturing gradient gel electrophoresis (DGGE), denaturing high performance liquid chromatography (DHPLC), and direct sequencing. In recent years, it has been shown that a significant proportion of undetermined alleles consist of large deletions overlapping one or more exons. These deletions have been difficult to detect in compound heterozygotes using gene-scanning methods due to a masking effect of the non-deleted allele. To date, no systematic search has been carried out for such exon deletions in Italian patients with phenylketonuria or mild hyperphenylalaninemia. We used multiplex ligation-dependent probe amplification (MLPA), comparative multiplex dosage analysis (CMDA), and real-time PCR to search for both large deletions and duplications of the phenylalanine hydroxylase gene in Italian hyperphenylalaninemia patients. Four deletions removing different phenylalanine hydroxylase (PAH) gene exons were identified in 12 patients. Two of these deletions involving exons 4-5-6-7-8 (systematic name c.353-?_912 + ?del) and exon 6 (systematic name c.510-?_706 + ?del) have not been reported previously. In this study, we show that exon deletion of the PAH gene accounts for 1.7% of all mutant PAH alleles in Italian hyperphenylalaninemics.

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Section: Methods Patientsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Exon deletions of the phenylalanine hydroxylase gene in Italian hyperphenylalaninemics

et al. 2010

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“…Table 1 provides overlapping data for the northern region of Piemont [Giannattasio et al, 2001], northern and central Italy combined [Dianzani et al, 1994], three southern regions combined (Calabria, Campania, Puglia/Basilicata) [Giannattasio et al, 2001], and data from Sicily [Mirisola et al, 2001]. There are relatively high frequencies of R252W, IVS12+1G4A, IVS7+1G4A, and R158Q in Northern Italy, while IVS10-11G4A and L48S are quite common in the south.…”

Section: Italy Spain and Portugalmentioning

confidence: 99%

“…Calculation of homozygosity at the PAH locus [Guldberg et al, 1996a;Scriver and Kaufman, 2001] has been used to distinguish relatively homogeneous populations (such as Yemenite Jews or the people of Iceland) from those that have experienced immigration from many regions in the past (such as the populations of Sicily, Germany, or the United States). The remarkable heterogeneity of PKU in Sicily [Guldberg et al, 1993b;Mirisola et al, 2001] thus reflects millennia of migration in the Mediterranean. Again however, such data have to be used with caution: the high homozygosity at the PAH locus in eastern Europe is caused by the prevalence of a single mutation in a large region and not necessarily by lack of migration.…”

Section: Pku Mutations and The History Of European Peoplesmentioning

confidence: 99%

Phenylketonuria mutations in Europe

2003

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For the PKU Special IssuePhenylketonuria (PKU) is heterogeneous. More than 400 different mutations in the phenylalanine hydroxylase (PAH) gene have been identified. In a systematic review of the molecular genetics of PKU in Europe we identified 29 mutations that may be regarded as prevalent in European populations.Comprehensive regional data for these mutations were collated from all available studies. The spectrum of mutations found in individual regions results from a combination of factors including founder effect, range expansion and migration, genetic drift, and probably heterozygote advantage.

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“…For each transfection, PAH activity was assayed on 50 µg of proteins, in duplicate, as previously reported [24]. This test measures the amount of 14 C radiolabeled Phe converted to…”

Section: Enzyme Analysismentioning

confidence: 99%

Five human phenylalanine hydroxylase proteins identified in mild hyperphenylalaninemia patients are disease-causing variants

Daniele

Cardillo²,

Pennino³

et al. 2008

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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Hyperphenylalaninemia is a group of autosomal recessive disorders caused by a wide range of phenylalanine hydroxylase (PAH) gene variants. To study the effects of mutations on PAH activity, we have reproduced five mutations (p.N223Y, p.R297L, p.F382L, p.K398N and p.Q419R) that we recently identified in a population of Southern Italy. Transient expression of mutant full-length cDNAs in human HEK293 cells yielded PAH variants whose l-phenylalanine hydroxylase activity was between 40% and 70% that of the wild-type enzyme. Moreover, Western blot analysis revealed a 50-kD monomer in all mutants thereby indicating normal synthesis of the mutant proteins. Because of the clinical mild nature of the phenotypes we performed an in vivo BH4 loading test. This was positive in all tested patients, which indicates that they are likely to respond to the coenzyme in vivo. We also analysed the environment of each mutation site in the available crystal structures of PAH by using molecular graphics tools. The structural alteration produced by each mutation was elucidated and correlated to the mutated properties of the mutant enzymes. All the data obtained demonstrate the disease-causing nature of the five novel variants.

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PAH Gene Mutations in the Sicilian Population: Association with Minihaplotypes and Expression Analysis

Cited by 15 publications

References 28 publications

Exon deletions of the phenylalanine hydroxylase gene in Italian hyperphenylalaninemics

Exon deletions of the phenylalanine hydroxylase gene in Italian hyperphenylalaninemics

Phenylketonuria mutations in Europe

Five human phenylalanine hydroxylase proteins identified in mild hyperphenylalaninemia patients are disease-causing variants

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