splitGFP Technology Reveals Dose-Dependent ER-Mitochondria Interface Modulation by α-Synuclein A53T and A30P Mutants
Familial Parkinson’s disease (PD) is associated with duplication or mutations of α-synuclein gene, whose product is a presynaptic cytosolic protein also found in mitochondria and in mitochondrial-associated ER membranes. We have originally shown the role of α-syn as a mod...
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MDPI AG
2019-09-01
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| Online Access: | https://www.mdpi.com/2073-4409/8/9/1072 |
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| author | Tito Calì Denis Ottolini Mattia Vicario Cristina Catoni Francesca Vallese Domenico Cieri Lucia Barazzuol Marisa Brini |
| author_facet | Tito Calì Denis Ottolini Mattia Vicario Cristina Catoni Francesca Vallese Domenico Cieri Lucia Barazzuol Marisa Brini |
| author_sort | Tito Calì |
| collection | DOAJ |
| description | Familial Parkinson’s disease (PD) is associated with duplication or mutations of α-synuclein gene, whose product is a presynaptic cytosolic protein also found in mitochondria and in mitochondrial-associated ER membranes. We have originally shown the role of α-syn as a modulator of the ER-mitochondria interface and mitochondrial Ca<sup>2+</sup> transients, suggesting that, at mild levels of expression, α-syn sustains cell metabolism. Here, we investigated the possibility that α-syn action on ER-mitochondria tethering could be compromised by the presence of PD-related mutations. The clarification of this aspect could contribute to elucidate key mechanisms underlying PD. The findings reported so far are not consistent, possibly because of the different methods used to evaluate ER-mitochondria connectivity. Here, the effects of the PD-related α-syn mutations A53T and A30P on ER-mitochondria relationship were investigated in respect to Ca<sup>2+</sup> handling and mitochondrial function using a newly generated SPLICS sensor and aequorin-based Ca<sup>2+</sup>measurements. We provided evidence that A53T and A30P amino acid substitution does not affect the ability of α-syn to enhance ER/mitochondria tethering and mitochondrial Ca<sup>2+</sup> transients, but that this action was lost as soon as a high amount of TAT-delivered A53T and A30P α-syn mutants caused the redistribution of α-syn from cytoplasm to foci. Our results suggest a loss of function mechanism and highlight a possible connection between α-syn and ER-mitochondria Ca<sup>2+</sup> cross-talk impairment to the pathogenesis of PD. |
| first_indexed | 2024-03-12T09:08:49Z |
| format | Article |
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| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-12T09:08:49Z |
| publishDate | 2019-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-49f562495f8c46f4bb2b4a96eda9f7e62023-09-02T15:04:48ZengMDPI AGCells2073-44092019-09-0189107210.3390/cells8091072cells8091072splitGFP Technology Reveals Dose-Dependent ER-Mitochondria Interface Modulation by α-Synuclein A53T and A30P MutantsTito Calì0Denis Ottolini1Mattia Vicario2Cristina Catoni3Francesca Vallese4Domenico Cieri5Lucia Barazzuol6Marisa Brini7Department of Biomedical Sciences, University of Padova, Padova 35131, ItalyDepartment of Biology, University of Padova, Padova 35131, ItalyDepartment of Biomedical Sciences, University of Padova, Padova 35131, ItalyDepartment of Biology, University of Padova, Padova 35131, ItalyDepartment of Biomedical Sciences, University of Padova, Padova 35131, ItalyDepartment of Biomedical Sciences, University of Padova, Padova 35131, ItalyDepartment of Biomedical Sciences, University of Padova, Padova 35131, ItalyDepartment of Biology, University of Padova, Padova 35131, ItalyFamilial Parkinson’s disease (PD) is associated with duplication or mutations of α-synuclein gene, whose product is a presynaptic cytosolic protein also found in mitochondria and in mitochondrial-associated ER membranes. We have originally shown the role of α-syn as a modulator of the ER-mitochondria interface and mitochondrial Ca<sup>2+</sup> transients, suggesting that, at mild levels of expression, α-syn sustains cell metabolism. Here, we investigated the possibility that α-syn action on ER-mitochondria tethering could be compromised by the presence of PD-related mutations. The clarification of this aspect could contribute to elucidate key mechanisms underlying PD. The findings reported so far are not consistent, possibly because of the different methods used to evaluate ER-mitochondria connectivity. Here, the effects of the PD-related α-syn mutations A53T and A30P on ER-mitochondria relationship were investigated in respect to Ca<sup>2+</sup> handling and mitochondrial function using a newly generated SPLICS sensor and aequorin-based Ca<sup>2+</sup>measurements. We provided evidence that A53T and A30P amino acid substitution does not affect the ability of α-syn to enhance ER/mitochondria tethering and mitochondrial Ca<sup>2+</sup> transients, but that this action was lost as soon as a high amount of TAT-delivered A53T and A30P α-syn mutants caused the redistribution of α-syn from cytoplasm to foci. Our results suggest a loss of function mechanism and highlight a possible connection between α-syn and ER-mitochondria Ca<sup>2+</sup> cross-talk impairment to the pathogenesis of PD.https://www.mdpi.com/2073-4409/8/9/1072Parkinson’s diseasealpha-synucleincalciummitochondriaER-mitochondria contact sites |
| spellingShingle | Tito Calì Denis Ottolini Mattia Vicario Cristina Catoni Francesca Vallese Domenico Cieri Lucia Barazzuol Marisa Brini splitGFP Technology Reveals Dose-Dependent ER-Mitochondria Interface Modulation by α-Synuclein A53T and A30P Mutants Cells Parkinson’s disease alpha-synuclein calcium mitochondria ER-mitochondria contact sites |
| title | splitGFP Technology Reveals Dose-Dependent ER-Mitochondria Interface Modulation by α-Synuclein A53T and A30P Mutants |
| title_full | splitGFP Technology Reveals Dose-Dependent ER-Mitochondria Interface Modulation by α-Synuclein A53T and A30P Mutants |
| title_fullStr | splitGFP Technology Reveals Dose-Dependent ER-Mitochondria Interface Modulation by α-Synuclein A53T and A30P Mutants |
| title_full_unstemmed | splitGFP Technology Reveals Dose-Dependent ER-Mitochondria Interface Modulation by α-Synuclein A53T and A30P Mutants |
| title_short | splitGFP Technology Reveals Dose-Dependent ER-Mitochondria Interface Modulation by α-Synuclein A53T and A30P Mutants |
| title_sort | splitgfp technology reveals dose dependent er mitochondria interface modulation by α synuclein a53t and a30p mutants |
| topic | Parkinson’s disease alpha-synuclein calcium mitochondria ER-mitochondria contact sites |
| url | https://www.mdpi.com/2073-4409/8/9/1072 |
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