New mechanistic insights into the RAS-SIN1 interaction at the membrane
Stress-activated MAP kinase-interacting protein 1 (SIN1) is a central member of the mTORC2 complex that contains an N-terminal domain (NTD), a conserved region in the middle (CRIM), a RAS-binding domain (RBD), and a pleckstrin homology domain. Recent studies provided valuable structural and function...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-10-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2022.987754/full |
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| author | Silke Pudewell Jana Lissy Hossein Nakhaeizadeh Niloufar Mosaddeghzadeh Saeideh Nakhaei-Rad Saeideh Nakhaei-Rad Radovan Dvorsky Radovan Dvorsky Mohammad R. Ahmadian |
| author_facet | Silke Pudewell Jana Lissy Hossein Nakhaeizadeh Niloufar Mosaddeghzadeh Saeideh Nakhaei-Rad Saeideh Nakhaei-Rad Radovan Dvorsky Radovan Dvorsky Mohammad R. Ahmadian |
| author_sort | Silke Pudewell |
| collection | DOAJ |
| description | Stress-activated MAP kinase-interacting protein 1 (SIN1) is a central member of the mTORC2 complex that contains an N-terminal domain (NTD), a conserved region in the middle (CRIM), a RAS-binding domain (RBD), and a pleckstrin homology domain. Recent studies provided valuable structural and functional insights into the interactions of SIN1 and the RAS-binding domain of RAS proteins. However, the mechanism for a reciprocal interaction of the RBD-PH tandem with RAS proteins and the membrane as an upstream event to spatiotemporal mTORC2 regulation is not clear. The biochemical assays in this study led to the following results: 1) all classical RAS paralogs, including HRAS, KRAS4A, KRAS4B, and NRAS, can bind to SIN1-RBD in biophysical and SIN1 full length (FL) in cell biology experiments; 2) the SIN1-PH domain modulates interactions with various types of membrane phosphoinositides and constantly maintains a pool of SIN1 at the membrane; and 3) a KRAS4A-dependent decrease in membrane binding of the SIN1-RBD-PH tandem was observed, suggesting for the first time a mechanistic influence of KRAS4A on SIN1 membrane association. Our study strengthens the current mechanistic understanding of SIN1-RAS interaction and suggests membrane interaction as a key event in the control of mTORC2-dependent and mTORC2-independent SIN1 function. |
| first_indexed | 2024-04-12T00:56:42Z |
| format | Article |
| id | doaj.art-5d40e5ad25814d9da32bd2c0caba6148 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-04-12T00:56:42Z |
| publishDate | 2022-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-5d40e5ad25814d9da32bd2c0caba61482022-12-22T03:54:35ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2022-10-011010.3389/fcell.2022.987754987754New mechanistic insights into the RAS-SIN1 interaction at the membraneSilke Pudewell0Jana Lissy1Hossein Nakhaeizadeh2Niloufar Mosaddeghzadeh3Saeideh Nakhaei-Rad4Saeideh Nakhaei-Rad5Radovan Dvorsky6Radovan Dvorsky7Mohammad R. Ahmadian8Institute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, GermanyStem Cell Biology and Regenerative Medicine Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, IranInstitute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, GermanyCenter for Interdisciplinary Biosciences, P. J. Šafárik University, Košice, SlovakiaInstitute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, GermanyStress-activated MAP kinase-interacting protein 1 (SIN1) is a central member of the mTORC2 complex that contains an N-terminal domain (NTD), a conserved region in the middle (CRIM), a RAS-binding domain (RBD), and a pleckstrin homology domain. Recent studies provided valuable structural and functional insights into the interactions of SIN1 and the RAS-binding domain of RAS proteins. However, the mechanism for a reciprocal interaction of the RBD-PH tandem with RAS proteins and the membrane as an upstream event to spatiotemporal mTORC2 regulation is not clear. The biochemical assays in this study led to the following results: 1) all classical RAS paralogs, including HRAS, KRAS4A, KRAS4B, and NRAS, can bind to SIN1-RBD in biophysical and SIN1 full length (FL) in cell biology experiments; 2) the SIN1-PH domain modulates interactions with various types of membrane phosphoinositides and constantly maintains a pool of SIN1 at the membrane; and 3) a KRAS4A-dependent decrease in membrane binding of the SIN1-RBD-PH tandem was observed, suggesting for the first time a mechanistic influence of KRAS4A on SIN1 membrane association. Our study strengthens the current mechanistic understanding of SIN1-RAS interaction and suggests membrane interaction as a key event in the control of mTORC2-dependent and mTORC2-independent SIN1 function.https://www.frontiersin.org/articles/10.3389/fcell.2022.987754/fullRASRAS familySIN1MAPKAP1mTORC2ras binding domain |
| spellingShingle | Silke Pudewell Jana Lissy Hossein Nakhaeizadeh Niloufar Mosaddeghzadeh Saeideh Nakhaei-Rad Saeideh Nakhaei-Rad Radovan Dvorsky Radovan Dvorsky Mohammad R. Ahmadian New mechanistic insights into the RAS-SIN1 interaction at the membrane Frontiers in Cell and Developmental Biology RAS RAS family SIN1 MAPKAP1 mTORC2 ras binding domain |
| title | New mechanistic insights into the RAS-SIN1 interaction at the membrane |
| title_full | New mechanistic insights into the RAS-SIN1 interaction at the membrane |
| title_fullStr | New mechanistic insights into the RAS-SIN1 interaction at the membrane |
| title_full_unstemmed | New mechanistic insights into the RAS-SIN1 interaction at the membrane |
| title_short | New mechanistic insights into the RAS-SIN1 interaction at the membrane |
| title_sort | new mechanistic insights into the ras sin1 interaction at the membrane |
| topic | RAS RAS family SIN1 MAPKAP1 mTORC2 ras binding domain |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2022.987754/full |
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