Molecular basis for differential activation of p101 and p84 complexes of PI3Kγ by Ras and GPCRs
Summary: Class IB phosphoinositide 3-kinase (PI3Kγ) is activated in immune cells and can form two distinct complexes (p110γ-p84 and p110γ-p101), which are differentially activated by G protein-coupled receptors (GPCRs) and Ras. Using a combination of X-ray crystallography, hydrogen deuterium exchang...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | Cell Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124723001833 |
| _version_ | 1797894414690942976 |
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| author | Manoj K. Rathinaswamy Meredith L. Jenkins Benjamin R. Duewell Xuxiao Zhang Noah J. Harris John T. Evans Jordan T.B. Stariha Udit Dalwadi Kaelin D. Fleming Harish Ranga-Prasad Calvin K. Yip Roger L. Williams Scott D. Hansen John E. Burke |
| author_facet | Manoj K. Rathinaswamy Meredith L. Jenkins Benjamin R. Duewell Xuxiao Zhang Noah J. Harris John T. Evans Jordan T.B. Stariha Udit Dalwadi Kaelin D. Fleming Harish Ranga-Prasad Calvin K. Yip Roger L. Williams Scott D. Hansen John E. Burke |
| author_sort | Manoj K. Rathinaswamy |
| collection | DOAJ |
| description | Summary: Class IB phosphoinositide 3-kinase (PI3Kγ) is activated in immune cells and can form two distinct complexes (p110γ-p84 and p110γ-p101), which are differentially activated by G protein-coupled receptors (GPCRs) and Ras. Using a combination of X-ray crystallography, hydrogen deuterium exchange mass spectrometry (HDX-MS), electron microscopy, molecular modeling, single-molecule imaging, and activity assays, we identify molecular differences between p110γ-p84 and p110γ-p101 that explain their differential membrane recruitment and activation by Ras and GPCRs. The p110γ-p84 complex is dynamic compared with p110γ-p101. While p110γ-p101 is robustly recruited by Gβγ subunits, p110γ-p84 is weakly recruited to membranes by Gβγ subunits alone and requires recruitment by Ras to allow for Gβγ activation. We mapped two distinct Gβγ interfaces on p101 and the p110γ helical domain, with differences in the C-terminal domain of p84 and p101 conferring sensitivity of p110γ-p101 to Gβγ activation. Overall, our work provides key insight into the molecular basis for how PI3Kγ complexes are activated. |
| first_indexed | 2024-04-10T07:08:40Z |
| format | Article |
| id | doaj.art-adaec01d7c824f20bc3e5572c80805c4 |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-04-10T07:08:40Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-adaec01d7c824f20bc3e5572c80805c42023-02-27T04:07:07ZengElsevierCell Reports2211-12472023-03-01423112172Molecular basis for differential activation of p101 and p84 complexes of PI3Kγ by Ras and GPCRsManoj K. Rathinaswamy0Meredith L. Jenkins1Benjamin R. Duewell2Xuxiao Zhang3Noah J. Harris4John T. Evans5Jordan T.B. Stariha6Udit Dalwadi7Kaelin D. Fleming8Harish Ranga-Prasad9Calvin K. Yip10Roger L. Williams11Scott D. Hansen12John E. Burke13Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8W 2Y2, CanadaDepartment of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8W 2Y2, CanadaDepartment of Chemistry and Biochemistry, University of Oregon, Eugene, OR 97403, USA; Institute of Molecular Biology, University of Oregon, Eugene, OR 97403, USAMRC Laboratory of Molecular Biology, Cambridge, UKDepartment of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8W 2Y2, CanadaDepartment of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8W 2Y2, CanadaDepartment of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8W 2Y2, CanadaDepartment of Biochemistry and Molecular Biology, The University of British Columbia, Vancouver, BC V6T 1Z3, CanadaDepartment of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8W 2Y2, CanadaDepartment of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8W 2Y2, CanadaDepartment of Biochemistry and Molecular Biology, The University of British Columbia, Vancouver, BC V6T 1Z3, CanadaMRC Laboratory of Molecular Biology, Cambridge, UKDepartment of Chemistry and Biochemistry, University of Oregon, Eugene, OR 97403, USA; Institute of Molecular Biology, University of Oregon, Eugene, OR 97403, USA; Corresponding authorDepartment of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8W 2Y2, Canada; Department of Biochemistry and Molecular Biology, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada; Corresponding authorSummary: Class IB phosphoinositide 3-kinase (PI3Kγ) is activated in immune cells and can form two distinct complexes (p110γ-p84 and p110γ-p101), which are differentially activated by G protein-coupled receptors (GPCRs) and Ras. Using a combination of X-ray crystallography, hydrogen deuterium exchange mass spectrometry (HDX-MS), electron microscopy, molecular modeling, single-molecule imaging, and activity assays, we identify molecular differences between p110γ-p84 and p110γ-p101 that explain their differential membrane recruitment and activation by Ras and GPCRs. The p110γ-p84 complex is dynamic compared with p110γ-p101. While p110γ-p101 is robustly recruited by Gβγ subunits, p110γ-p84 is weakly recruited to membranes by Gβγ subunits alone and requires recruitment by Ras to allow for Gβγ activation. We mapped two distinct Gβγ interfaces on p101 and the p110γ helical domain, with differences in the C-terminal domain of p84 and p101 conferring sensitivity of p110γ-p101 to Gβγ activation. Overall, our work provides key insight into the molecular basis for how PI3Kγ complexes are activated.http://www.sciencedirect.com/science/article/pii/S2211124723001833CP: Cell biology |
| spellingShingle | Manoj K. Rathinaswamy Meredith L. Jenkins Benjamin R. Duewell Xuxiao Zhang Noah J. Harris John T. Evans Jordan T.B. Stariha Udit Dalwadi Kaelin D. Fleming Harish Ranga-Prasad Calvin K. Yip Roger L. Williams Scott D. Hansen John E. Burke Molecular basis for differential activation of p101 and p84 complexes of PI3Kγ by Ras and GPCRs Cell Reports CP: Cell biology |
| title | Molecular basis for differential activation of p101 and p84 complexes of PI3Kγ by Ras and GPCRs |
| title_full | Molecular basis for differential activation of p101 and p84 complexes of PI3Kγ by Ras and GPCRs |
| title_fullStr | Molecular basis for differential activation of p101 and p84 complexes of PI3Kγ by Ras and GPCRs |
| title_full_unstemmed | Molecular basis for differential activation of p101 and p84 complexes of PI3Kγ by Ras and GPCRs |
| title_short | Molecular basis for differential activation of p101 and p84 complexes of PI3Kγ by Ras and GPCRs |
| title_sort | molecular basis for differential activation of p101 and p84 complexes of pi3kγ by ras and gpcrs |
| topic | CP: Cell biology |
| url | http://www.sciencedirect.com/science/article/pii/S2211124723001833 |
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