Topology and Function of the <i>S. cerevisiae</i> Autophagy Protein Atg15
The putative phospholipase Atg15 is required for the intravacuolar lysis of autophagic bodies and MVB vesicles. Intracellular membrane lysis is a highly sophisticated mechanism that is not fully understood. The amino-terminal transmembrane domain of Atg15 contains the sorting signal for entry into t...
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MDPI AG
2023-08-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/12/16/2056 |
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| author | Lisa Marquardt Marco Montino Yvonne Mühe Petra Schlotterhose Michael Thumm |
| author_facet | Lisa Marquardt Marco Montino Yvonne Mühe Petra Schlotterhose Michael Thumm |
| author_sort | Lisa Marquardt |
| collection | DOAJ |
| description | The putative phospholipase Atg15 is required for the intravacuolar lysis of autophagic bodies and MVB vesicles. Intracellular membrane lysis is a highly sophisticated mechanism that is not fully understood. The amino-terminal transmembrane domain of Atg15 contains the sorting signal for entry into the MVB pathway. By replacing this domain, we generated chimeras located in the cytosol, the vacuole membrane, and the lumen. The variants at the vacuole membrane and in the lumen were highly active. Together with the absence of Atg15 from the phagophore and autophagic bodies, this suggests that, within the vacuole, Atg15 can lyse vesicles where it is not embedded. In-depth topological analyses showed that Atg15 is a single membrane-spanning protein with the amino-terminus in the cytosol and the rest, including the active site motif, in the ER lumen. Remarkably, only membrane-embedded Atg15 variants affected growth when overexpressed. The growth defects depended on its active site serine 332, showing that it was linked to the enzymatic activity of Atg15. Interestingly, the growth defects were independent of vacuolar proteinase A and vacuolar acidification. |
| first_indexed | 2024-03-11T00:03:35Z |
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| id | doaj.art-a0c41cb0a4cc4df2bfc03180570bdbf1 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T00:03:35Z |
| publishDate | 2023-08-01 |
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| spelling | doaj.art-a0c41cb0a4cc4df2bfc03180570bdbf12023-11-19T00:36:47ZengMDPI AGCells2073-44092023-08-011216205610.3390/cells12162056Topology and Function of the <i>S. cerevisiae</i> Autophagy Protein Atg15Lisa Marquardt0Marco Montino1Yvonne Mühe2Petra Schlotterhose3Michael Thumm4Institute of Cellular Biochemistry, University Medicine, Humboldtallee 23, D-37073 Goettingen, GermanyInstitute of Cellular Biochemistry, University Medicine, Humboldtallee 23, D-37073 Goettingen, GermanyInstitute of Cellular Biochemistry, University Medicine, Humboldtallee 23, D-37073 Goettingen, GermanyInstitute of Cellular Biochemistry, University Medicine, Humboldtallee 23, D-37073 Goettingen, GermanyInstitute of Cellular Biochemistry, University Medicine, Humboldtallee 23, D-37073 Goettingen, GermanyThe putative phospholipase Atg15 is required for the intravacuolar lysis of autophagic bodies and MVB vesicles. Intracellular membrane lysis is a highly sophisticated mechanism that is not fully understood. The amino-terminal transmembrane domain of Atg15 contains the sorting signal for entry into the MVB pathway. By replacing this domain, we generated chimeras located in the cytosol, the vacuole membrane, and the lumen. The variants at the vacuole membrane and in the lumen were highly active. Together with the absence of Atg15 from the phagophore and autophagic bodies, this suggests that, within the vacuole, Atg15 can lyse vesicles where it is not embedded. In-depth topological analyses showed that Atg15 is a single membrane-spanning protein with the amino-terminus in the cytosol and the rest, including the active site motif, in the ER lumen. Remarkably, only membrane-embedded Atg15 variants affected growth when overexpressed. The growth defects depended on its active site serine 332, showing that it was linked to the enzymatic activity of Atg15. Interestingly, the growth defects were independent of vacuolar proteinase A and vacuolar acidification.https://www.mdpi.com/2073-4409/12/16/2056autophagic bodylysismacroautophagymicroautophagyyeastphospholipase |
| spellingShingle | Lisa Marquardt Marco Montino Yvonne Mühe Petra Schlotterhose Michael Thumm Topology and Function of the <i>S. cerevisiae</i> Autophagy Protein Atg15 Cells autophagic body lysis macroautophagy microautophagy yeast phospholipase |
| title | Topology and Function of the <i>S. cerevisiae</i> Autophagy Protein Atg15 |
| title_full | Topology and Function of the <i>S. cerevisiae</i> Autophagy Protein Atg15 |
| title_fullStr | Topology and Function of the <i>S. cerevisiae</i> Autophagy Protein Atg15 |
| title_full_unstemmed | Topology and Function of the <i>S. cerevisiae</i> Autophagy Protein Atg15 |
| title_short | Topology and Function of the <i>S. cerevisiae</i> Autophagy Protein Atg15 |
| title_sort | topology and function of the i s cerevisiae i autophagy protein atg15 |
| topic | autophagic body lysis macroautophagy microautophagy yeast phospholipase |
| url | https://www.mdpi.com/2073-4409/12/16/2056 |
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