Structural basis for the interaction between the Drosophila RTK Sevenless (dROS1) and the GPCR BOSS
Abstract Sevenless, the Drosophila homologue of ROS1 (University of Rochester Sarcoma) (herein, dROS1) is a receptor tyrosine kinase (RTK) essential for the differentiation of Drosophila R7 photoreceptor cells. Activation of dROS1 is mediated by binding to the extracellular region (ECR) of the GPCR...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-01-01
|
| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-55943-6 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832594534348881920 |
|---|---|
| author | Jianan Zhang Yuko Tsutsui Hengyi Li Tongqing Li Yueyue Wang Salma Laraki Sofia Alarcon-Frias Steven E. Stayrook Daryl E. Klein |
| author_facet | Jianan Zhang Yuko Tsutsui Hengyi Li Tongqing Li Yueyue Wang Salma Laraki Sofia Alarcon-Frias Steven E. Stayrook Daryl E. Klein |
| author_sort | Jianan Zhang |
| collection | DOAJ |
| description | Abstract Sevenless, the Drosophila homologue of ROS1 (University of Rochester Sarcoma) (herein, dROS1) is a receptor tyrosine kinase (RTK) essential for the differentiation of Drosophila R7 photoreceptor cells. Activation of dROS1 is mediated by binding to the extracellular region (ECR) of the GPCR (G protein coupled receptor) BOSS (Bride Of Sevenless) on adjacent cells. Activation of dROS1 by BOSS leads to subsequent downstream signaling pathways including SOS (Son of Sevenless). However, the physical basis for how dROS1 interacts with BOSS has long remained unknown. Here we provide a cryo-EM structure of dROS1’s extracellular region, which mediates ligand binding. We show that the extracellular region of dROS1 adopts a folded-over conformation stabilized by an N-terminal domain comprised of two disulfide stapled helical hairpins. We further narrowed down the interacting binding epitopes on both dROS1 and BOSS using hydrogen-deuterium exchange mass spectrometry (HDX-MS). This includes beta-strands in dROS1’s third Fibronectin type III (FNIII) domain and a C-terminal peptide in BOSS’ ECR. Our mutagenesis studies, coupled with AlphaFold complex predictions, support a binding interaction mediated by a hydrophobic interaction and beta-strand augmentation between these regions. Our findings provide a fundamental understanding of the regulatory function of dROS1 and further provide mechanistic insight into the human ortholog and oncogene ROS1. |
| format | Article |
| id | doaj-art-22bf27c2c889410fa8be600c69e67f7a |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-22bf27c2c889410fa8be600c69e67f7a2025-01-19T12:30:31ZengNature PortfolioNature Communications2041-17232025-01-0116111210.1038/s41467-025-55943-6Structural basis for the interaction between the Drosophila RTK Sevenless (dROS1) and the GPCR BOSSJianan Zhang0Yuko Tsutsui1Hengyi Li2Tongqing Li3Yueyue Wang4Salma Laraki5Sofia Alarcon-Frias6Steven E. Stayrook7Daryl E. Klein8Department of Pharmacology, Yale University School of MedicineDepartment of Pharmacology, Yale University School of MedicineDepartment of Pharmacology, Yale University School of MedicineDepartment of Pharmacology, Yale University School of MedicineYale Cancer Biology Institute, Yale UniversityDepartment of Pharmacology, Yale University School of MedicineDepartment of Pharmacology, Yale University School of MedicineDepartment of Pharmacology, Yale University School of MedicineDepartment of Pharmacology, Yale University School of MedicineAbstract Sevenless, the Drosophila homologue of ROS1 (University of Rochester Sarcoma) (herein, dROS1) is a receptor tyrosine kinase (RTK) essential for the differentiation of Drosophila R7 photoreceptor cells. Activation of dROS1 is mediated by binding to the extracellular region (ECR) of the GPCR (G protein coupled receptor) BOSS (Bride Of Sevenless) on adjacent cells. Activation of dROS1 by BOSS leads to subsequent downstream signaling pathways including SOS (Son of Sevenless). However, the physical basis for how dROS1 interacts with BOSS has long remained unknown. Here we provide a cryo-EM structure of dROS1’s extracellular region, which mediates ligand binding. We show that the extracellular region of dROS1 adopts a folded-over conformation stabilized by an N-terminal domain comprised of two disulfide stapled helical hairpins. We further narrowed down the interacting binding epitopes on both dROS1 and BOSS using hydrogen-deuterium exchange mass spectrometry (HDX-MS). This includes beta-strands in dROS1’s third Fibronectin type III (FNIII) domain and a C-terminal peptide in BOSS’ ECR. Our mutagenesis studies, coupled with AlphaFold complex predictions, support a binding interaction mediated by a hydrophobic interaction and beta-strand augmentation between these regions. Our findings provide a fundamental understanding of the regulatory function of dROS1 and further provide mechanistic insight into the human ortholog and oncogene ROS1.https://doi.org/10.1038/s41467-025-55943-6 |
| spellingShingle | Jianan Zhang Yuko Tsutsui Hengyi Li Tongqing Li Yueyue Wang Salma Laraki Sofia Alarcon-Frias Steven E. Stayrook Daryl E. Klein Structural basis for the interaction between the Drosophila RTK Sevenless (dROS1) and the GPCR BOSS Nature Communications |
| title | Structural basis for the interaction between the Drosophila RTK Sevenless (dROS1) and the GPCR BOSS |
| title_full | Structural basis for the interaction between the Drosophila RTK Sevenless (dROS1) and the GPCR BOSS |
| title_fullStr | Structural basis for the interaction between the Drosophila RTK Sevenless (dROS1) and the GPCR BOSS |
| title_full_unstemmed | Structural basis for the interaction between the Drosophila RTK Sevenless (dROS1) and the GPCR BOSS |
| title_short | Structural basis for the interaction between the Drosophila RTK Sevenless (dROS1) and the GPCR BOSS |
| title_sort | structural basis for the interaction between the drosophila rtk sevenless dros1 and the gpcr boss |
| url | https://doi.org/10.1038/s41467-025-55943-6 |
| work_keys_str_mv | AT jiananzhang structuralbasisfortheinteractionbetweenthedrosophilartksevenlessdros1andthegpcrboss AT yukotsutsui structuralbasisfortheinteractionbetweenthedrosophilartksevenlessdros1andthegpcrboss AT hengyili structuralbasisfortheinteractionbetweenthedrosophilartksevenlessdros1andthegpcrboss AT tongqingli structuralbasisfortheinteractionbetweenthedrosophilartksevenlessdros1andthegpcrboss AT yueyuewang structuralbasisfortheinteractionbetweenthedrosophilartksevenlessdros1andthegpcrboss AT salmalaraki structuralbasisfortheinteractionbetweenthedrosophilartksevenlessdros1andthegpcrboss AT sofiaalarconfrias structuralbasisfortheinteractionbetweenthedrosophilartksevenlessdros1andthegpcrboss AT stevenestayrook structuralbasisfortheinteractionbetweenthedrosophilartksevenlessdros1andthegpcrboss AT daryleklein structuralbasisfortheinteractionbetweenthedrosophilartksevenlessdros1andthegpcrboss |