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Mutational analysis of Peroxiredoxin IV: exclusion of a positional candidate for multinodular goitre.

Giardina E, Capon F, D'Apice MR, Amati F, Arturi F, Filetti S, Bonifazi E, Pucci S, Conte C, Novelli G - BMC Med. Genet. (2002)

Bottom Line: No causative mutations were found.Finally, using RT-PCR from tissue-specific RNA, no differences of PRX-IV expression-levels were detected between affected and unaffected samples.Based on sequence and genomic analysis, Prx-IV is very unlikely to be the MNG2 gene.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biopathology, "Tor Vergata" University of Rome, Italy. emizago@yahoo.com

ABSTRACT

Background: Multinodular goitre (MNG) is a common disorder characterised by an enlargement of the thyroid, occurring as a compensatory response to hormonogenesis impairment. The incidence of MNG is dependent on sex (female:male ratio 5:1) and several reports have documented a genetic basis for the disease. Last year we mapped a MNG locus to chromosome Xp22 in a region containing the peroxiredoxin IV (Prx-IV) gene. Since Prx-IV is involved in the removal of H2O2 in thyroid cells, we hypothesize that mutations in Prx-IV gene are involved in pathogenesis of MNG.

Methods: Four individuals (2 affected, 2 unrelated unaffected) were sequenced using automated methods. All individuals were originated from the original three-generation Italian family described in previous studies. A Southern blot analysis using a Prx-IV full-length cDNA as a probe was performed in order to exclude genomic rearrangements and/or intronic mutations. In addition a RT-PCR of PRX-IV was performed in order to investigate expression alterations.

Results: No causative mutations were found. Two adjacent nucleotide substitutions were detected within introns 1 and 4. These changes were also detected in unaffected individuals, suggesting that they were innocuous polymorphisms. No gross genomic rearrangements and/or restriction fragment alterations were observed on Southern analysis. Finally, using RT-PCR from tissue-specific RNA, no differences of PRX-IV expression-levels were detected between affected and unaffected samples.

Conclusions: Based on sequence and genomic analysis, Prx-IV is very unlikely to be the MNG2 gene.

No MeSH data available.


Related in: MedlinePlus

Southern-blot analysis. Lane 1,3 : genomic DNA from a patient (L.F.) affected with Multinodular goitre digested respectively with EcoRI and HIND III. Lane 2,4 : genomic DNA from a normal control digested respectively with EcoRI and HIND III.
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Figure 1: Southern-blot analysis. Lane 1,3 : genomic DNA from a patient (L.F.) affected with Multinodular goitre digested respectively with EcoRI and HIND III. Lane 2,4 : genomic DNA from a normal control digested respectively with EcoRI and HIND III.

Mentions: Patient sequencing identified two adjacent substitutions in intron 1 (IVS1-9G>A and IVS1-10A>T), and one in intron 4 (IVS4+81T>C). All variants were also found in one unrelated unaffected male, thus ruling out the hypothesis that any of them might be disease-related. Coding region analysis failed to detect any additional substitutions and moreover promoter region analysis didn't detect other substitutions. Since the affected females carrying both alleles at intron 2 variants, the occurrence of a large gene deletion can be excluded. In addition, as shown in figure 1, no gross PRX-IV rearrangements were detected by Southern blot analysis.


Mutational analysis of Peroxiredoxin IV: exclusion of a positional candidate for multinodular goitre.

Giardina E, Capon F, D'Apice MR, Amati F, Arturi F, Filetti S, Bonifazi E, Pucci S, Conte C, Novelli G - BMC Med. Genet. (2002)

Southern-blot analysis. Lane 1,3 : genomic DNA from a patient (L.F.) affected with Multinodular goitre digested respectively with EcoRI and HIND III. Lane 2,4 : genomic DNA from a normal control digested respectively with EcoRI and HIND III.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC117784&req=5

Figure 1: Southern-blot analysis. Lane 1,3 : genomic DNA from a patient (L.F.) affected with Multinodular goitre digested respectively with EcoRI and HIND III. Lane 2,4 : genomic DNA from a normal control digested respectively with EcoRI and HIND III.
Mentions: Patient sequencing identified two adjacent substitutions in intron 1 (IVS1-9G>A and IVS1-10A>T), and one in intron 4 (IVS4+81T>C). All variants were also found in one unrelated unaffected male, thus ruling out the hypothesis that any of them might be disease-related. Coding region analysis failed to detect any additional substitutions and moreover promoter region analysis didn't detect other substitutions. Since the affected females carrying both alleles at intron 2 variants, the occurrence of a large gene deletion can be excluded. In addition, as shown in figure 1, no gross PRX-IV rearrangements were detected by Southern blot analysis.

Bottom Line: No causative mutations were found.Finally, using RT-PCR from tissue-specific RNA, no differences of PRX-IV expression-levels were detected between affected and unaffected samples.Based on sequence and genomic analysis, Prx-IV is very unlikely to be the MNG2 gene.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biopathology, "Tor Vergata" University of Rome, Italy. emizago@yahoo.com

ABSTRACT

Background: Multinodular goitre (MNG) is a common disorder characterised by an enlargement of the thyroid, occurring as a compensatory response to hormonogenesis impairment. The incidence of MNG is dependent on sex (female:male ratio 5:1) and several reports have documented a genetic basis for the disease. Last year we mapped a MNG locus to chromosome Xp22 in a region containing the peroxiredoxin IV (Prx-IV) gene. Since Prx-IV is involved in the removal of H2O2 in thyroid cells, we hypothesize that mutations in Prx-IV gene are involved in pathogenesis of MNG.

Methods: Four individuals (2 affected, 2 unrelated unaffected) were sequenced using automated methods. All individuals were originated from the original three-generation Italian family described in previous studies. A Southern blot analysis using a Prx-IV full-length cDNA as a probe was performed in order to exclude genomic rearrangements and/or intronic mutations. In addition a RT-PCR of PRX-IV was performed in order to investigate expression alterations.

Results: No causative mutations were found. Two adjacent nucleotide substitutions were detected within introns 1 and 4. These changes were also detected in unaffected individuals, suggesting that they were innocuous polymorphisms. No gross genomic rearrangements and/or restriction fragment alterations were observed on Southern analysis. Finally, using RT-PCR from tissue-specific RNA, no differences of PRX-IV expression-levels were detected between affected and unaffected samples.

Conclusions: Based on sequence and genomic analysis, Prx-IV is very unlikely to be the MNG2 gene.

No MeSH data available.


Related in: MedlinePlus