Sequence and copy number analyses of HEXB gene in patients affected by Sandhoff disease: functional characterization of 9 novel sequence variants.
Sandhoff disease (SD) is a lysosomal disorder caused by mutations in the HEXB gene. To date, 43 mutations of HEXB have been described, including 3 large deletions. Here, we have characterized 14 unrelated SD patients and developed a Multiplex Ligation-dependent Probe Amplification (MLPA) assay to in...
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0041516&type=printable |
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| author | Stefania Zampieri Silvia Cattarossi Ana Maria Oller Ramirez Camillo Rosano Charles Marques Lourenco Nadia Passon Isabella Moroni Graziella Uziel Antonella Pettinari Franco Stanzial Raquel Dodelson de Kremer Nydia Beatriz Azar Filiz Hazan Mirella Filocamo Bruno Bembi Andrea Dardis |
| author_facet | Stefania Zampieri Silvia Cattarossi Ana Maria Oller Ramirez Camillo Rosano Charles Marques Lourenco Nadia Passon Isabella Moroni Graziella Uziel Antonella Pettinari Franco Stanzial Raquel Dodelson de Kremer Nydia Beatriz Azar Filiz Hazan Mirella Filocamo Bruno Bembi Andrea Dardis |
| author_sort | Stefania Zampieri |
| collection | DOAJ |
| description | Sandhoff disease (SD) is a lysosomal disorder caused by mutations in the HEXB gene. To date, 43 mutations of HEXB have been described, including 3 large deletions. Here, we have characterized 14 unrelated SD patients and developed a Multiplex Ligation-dependent Probe Amplification (MLPA) assay to investigate the presence of large HEXB deletions. Overall, we identified 16 alleles, 9 of which were novel, including 4 sequence variation leading to aminoacid changes [c.626C>T (p.T209I), c.634C>A (p.H212N), c.926G>T (p.C309F), c.1451G>A (p.G484E)] 3 intronic mutations (c.1082+5G>A, c.1242+1G>A, c.1169+5G>A), 1 nonsense mutation c.146C>A (p.S49X) and 1 small in-frame deletion c.1260_1265delAGTTGA (p.V421_E422del). Using the new MLPA assay, 2 previously described deletions were identified. In vitro expression studies showed that proteins bearing aminoacid changes p.T209I and p.G484E presented a very low or absent activity, while proteins bearing the p.H212N and p.C309F changes retained a significant residual activity. The detrimental effect of the 3 novel intronic mutations on the HEXB mRNA processing was demonstrated using a minigene assay. Unprecedentedly, minigene studies revealed the presence of a novel alternative spliced HEXB mRNA variant also present in normal cells. In conclusion, we provided new insights into the molecular basis of SD and validated an MLPA assay for detecting large HEXB deletions. |
| format | Article |
| id | doaj-art-dd4476baf4464294933a6b2da0c8bb6e |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj-art-dd4476baf4464294933a6b2da0c8bb6e2025-08-20T02:30:51ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0177e4151610.1371/journal.pone.0041516Sequence and copy number analyses of HEXB gene in patients affected by Sandhoff disease: functional characterization of 9 novel sequence variants.Stefania ZampieriSilvia CattarossiAna Maria Oller RamirezCamillo RosanoCharles Marques LourencoNadia PassonIsabella MoroniGraziella UzielAntonella PettinariFranco StanzialRaquel Dodelson de KremerNydia Beatriz AzarFiliz HazanMirella FilocamoBruno BembiAndrea DardisSandhoff disease (SD) is a lysosomal disorder caused by mutations in the HEXB gene. To date, 43 mutations of HEXB have been described, including 3 large deletions. Here, we have characterized 14 unrelated SD patients and developed a Multiplex Ligation-dependent Probe Amplification (MLPA) assay to investigate the presence of large HEXB deletions. Overall, we identified 16 alleles, 9 of which were novel, including 4 sequence variation leading to aminoacid changes [c.626C>T (p.T209I), c.634C>A (p.H212N), c.926G>T (p.C309F), c.1451G>A (p.G484E)] 3 intronic mutations (c.1082+5G>A, c.1242+1G>A, c.1169+5G>A), 1 nonsense mutation c.146C>A (p.S49X) and 1 small in-frame deletion c.1260_1265delAGTTGA (p.V421_E422del). Using the new MLPA assay, 2 previously described deletions were identified. In vitro expression studies showed that proteins bearing aminoacid changes p.T209I and p.G484E presented a very low or absent activity, while proteins bearing the p.H212N and p.C309F changes retained a significant residual activity. The detrimental effect of the 3 novel intronic mutations on the HEXB mRNA processing was demonstrated using a minigene assay. Unprecedentedly, minigene studies revealed the presence of a novel alternative spliced HEXB mRNA variant also present in normal cells. In conclusion, we provided new insights into the molecular basis of SD and validated an MLPA assay for detecting large HEXB deletions.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0041516&type=printable |
| spellingShingle | Stefania Zampieri Silvia Cattarossi Ana Maria Oller Ramirez Camillo Rosano Charles Marques Lourenco Nadia Passon Isabella Moroni Graziella Uziel Antonella Pettinari Franco Stanzial Raquel Dodelson de Kremer Nydia Beatriz Azar Filiz Hazan Mirella Filocamo Bruno Bembi Andrea Dardis Sequence and copy number analyses of HEXB gene in patients affected by Sandhoff disease: functional characterization of 9 novel sequence variants. PLoS ONE |
| title | Sequence and copy number analyses of HEXB gene in patients affected by Sandhoff disease: functional characterization of 9 novel sequence variants. |
| title_full | Sequence and copy number analyses of HEXB gene in patients affected by Sandhoff disease: functional characterization of 9 novel sequence variants. |
| title_fullStr | Sequence and copy number analyses of HEXB gene in patients affected by Sandhoff disease: functional characterization of 9 novel sequence variants. |
| title_full_unstemmed | Sequence and copy number analyses of HEXB gene in patients affected by Sandhoff disease: functional characterization of 9 novel sequence variants. |
| title_short | Sequence and copy number analyses of HEXB gene in patients affected by Sandhoff disease: functional characterization of 9 novel sequence variants. |
| title_sort | sequence and copy number analyses of hexb gene in patients affected by sandhoff disease functional characterization of 9 novel sequence variants |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0041516&type=printable |
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