Pupylation‐Based Proximity Labeling Unravels a Comprehensive Protein and Phosphoprotein Interactome of the Arabidopsis TOR Complex
Abstract Target of rapamycin (TOR) is a signaling hub that integrates developmental, hormonal, and environmental signals to optimize carbon allocation and plant growth. In plant cells, TOR acts together with the proteins LST8‐1 and RAPTOR1 to form a core TOR complex (TORC). While these proteins comp...
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Wiley
2025-05-01
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| Series: | Advanced Science |
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| Online Access: | https://doi.org/10.1002/advs.202414496 |
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| author | Shuai Zheng Leonard Blaschek Delphine Pottier Luuk Robin Hoegen Dijkhof Beyza Özmen Peng Ken Lim Qiao Wen Tan Marek Mutwil Alexander Sebastian Hauser Staffan Persson |
| author_facet | Shuai Zheng Leonard Blaschek Delphine Pottier Luuk Robin Hoegen Dijkhof Beyza Özmen Peng Ken Lim Qiao Wen Tan Marek Mutwil Alexander Sebastian Hauser Staffan Persson |
| author_sort | Shuai Zheng |
| collection | DOAJ |
| description | Abstract Target of rapamycin (TOR) is a signaling hub that integrates developmental, hormonal, and environmental signals to optimize carbon allocation and plant growth. In plant cells, TOR acts together with the proteins LST8‐1 and RAPTOR1 to form a core TOR complex (TORC). While these proteins comprise a functional TORC, they engage with many other proteins to ensure precise signal outputs. Although TORC interactions have attracted significant attention in the recent past, large parts of the interactome are still unknown. In this resource study, PUP‐IT is adapted, a fully endogenously expressed protein proximity labeling toolbox, to map TORC protein–protein interactions using the core set of TORC as baits. It is outlined how this interactome is differentially phosphorylated during changes in carbon availability, uncovering putative direct TOR kinase targets. An AlphaFold‐Multimer approach is further used to validate many interactors, thus outlining a comprehensive TORC interactome that includes over a hundred new candidate interactors and provides an invaluable resource to the plant cell signaling community. |
| format | Article |
| id | doaj-art-6ddc69b23bd64279b15dfe14bb33bc6c |
| institution | DOAJ |
| issn | 2198-3844 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj-art-6ddc69b23bd64279b15dfe14bb33bc6c2025-08-20T03:13:30ZengWileyAdvanced Science2198-38442025-05-011219n/an/a10.1002/advs.202414496Pupylation‐Based Proximity Labeling Unravels a Comprehensive Protein and Phosphoprotein Interactome of the Arabidopsis TOR ComplexShuai Zheng0Leonard Blaschek1Delphine Pottier2Luuk Robin Hoegen Dijkhof3Beyza Özmen4Peng Ken Lim5Qiao Wen Tan6Marek Mutwil7Alexander Sebastian Hauser8Staffan Persson9Copenhagen Plant Science Center (CPSC) Department of Plant & Environmental Sciences University of Copenhagen Frederiksberg C 1871 DenmarkCopenhagen Plant Science Center (CPSC) Department of Plant & Environmental Sciences University of Copenhagen Frederiksberg C 1871 DenmarkCopenhagen Plant Science Center (CPSC) Department of Plant & Environmental Sciences University of Copenhagen Frederiksberg C 1871 DenmarkDepartment of Drug Design and Pharmacology Faculty of Health and Medical Sciences University of Copenhagen Copenhagen 2100 DenmarkCopenhagen Plant Science Center (CPSC) Department of Plant & Environmental Sciences University of Copenhagen Frederiksberg C 1871 DenmarkSchool of Biological Sciences Nanyang Technological University Singapore 637551 SingaporeSchool of Biological Sciences Nanyang Technological University Singapore 637551 SingaporeSchool of Biological Sciences Nanyang Technological University Singapore 637551 SingaporeDepartment of Drug Design and Pharmacology Faculty of Health and Medical Sciences University of Copenhagen Copenhagen 2100 DenmarkCopenhagen Plant Science Center (CPSC) Department of Plant & Environmental Sciences University of Copenhagen Frederiksberg C 1871 DenmarkAbstract Target of rapamycin (TOR) is a signaling hub that integrates developmental, hormonal, and environmental signals to optimize carbon allocation and plant growth. In plant cells, TOR acts together with the proteins LST8‐1 and RAPTOR1 to form a core TOR complex (TORC). While these proteins comprise a functional TORC, they engage with many other proteins to ensure precise signal outputs. Although TORC interactions have attracted significant attention in the recent past, large parts of the interactome are still unknown. In this resource study, PUP‐IT is adapted, a fully endogenously expressed protein proximity labeling toolbox, to map TORC protein–protein interactions using the core set of TORC as baits. It is outlined how this interactome is differentially phosphorylated during changes in carbon availability, uncovering putative direct TOR kinase targets. An AlphaFold‐Multimer approach is further used to validate many interactors, thus outlining a comprehensive TORC interactome that includes over a hundred new candidate interactors and provides an invaluable resource to the plant cell signaling community.https://doi.org/10.1002/advs.202414496AlphaFoldproximity labelingPUP‐ITsugar signalingtarget of rapamycin |
| spellingShingle | Shuai Zheng Leonard Blaschek Delphine Pottier Luuk Robin Hoegen Dijkhof Beyza Özmen Peng Ken Lim Qiao Wen Tan Marek Mutwil Alexander Sebastian Hauser Staffan Persson Pupylation‐Based Proximity Labeling Unravels a Comprehensive Protein and Phosphoprotein Interactome of the Arabidopsis TOR Complex Advanced Science AlphaFold proximity labeling PUP‐IT sugar signaling target of rapamycin |
| title | Pupylation‐Based Proximity Labeling Unravels a Comprehensive Protein and Phosphoprotein Interactome of the Arabidopsis TOR Complex |
| title_full | Pupylation‐Based Proximity Labeling Unravels a Comprehensive Protein and Phosphoprotein Interactome of the Arabidopsis TOR Complex |
| title_fullStr | Pupylation‐Based Proximity Labeling Unravels a Comprehensive Protein and Phosphoprotein Interactome of the Arabidopsis TOR Complex |
| title_full_unstemmed | Pupylation‐Based Proximity Labeling Unravels a Comprehensive Protein and Phosphoprotein Interactome of the Arabidopsis TOR Complex |
| title_short | Pupylation‐Based Proximity Labeling Unravels a Comprehensive Protein and Phosphoprotein Interactome of the Arabidopsis TOR Complex |
| title_sort | pupylation based proximity labeling unravels a comprehensive protein and phosphoprotein interactome of the arabidopsis tor complex |
| topic | AlphaFold proximity labeling PUP‐IT sugar signaling target of rapamycin |
| url | https://doi.org/10.1002/advs.202414496 |
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