Abstract
The cell-cell adhesion cadherin-catenin complexes recruit vinculin to the adherens junction (AJ) to modulate the mechanical couplings between neighboring cells. However, it is unclear how vinculin influences the AJ structure and function. Here, we identified two patches of salt bridges that lock vinculin in the head-tail autoinhibited conformation and reconstituted the full-length vinculin activation mimetics bound to the cadherin-catenin complex. The cadherin-catenin-vinculin complex contains multiple disordered linkers and is highly dynamic, which poses a challenge for structural studies. We determined the ensemble conformation of this complex using small-angle x-ray and selective deuteration/contrast variation small-angle neutron scattering. In the complex, both α-catenin and vinculin adopt an ensemble of flexible conformations, but vinculin has fully open conformations with the vinculin head and actin-binding tail domains well separated from each other. F-actin binding experiments show that the cadherin-catenin-vinculin complex binds and bundles F-actin. However, when the vinculin actin-binding domain is removed from the complex, only a minor fraction of the complex binds to F-actin. The results show that the dynamic cadherin-catenin-vinculin complex employs vinculin as the primary F-actin binding mode to strengthen AJ-cytoskeleton interactions.
| Original language | English |
|---|---|
| Pages (from-to) | 2456-2474 |
| Number of pages | 19 |
| Journal | Biophysical Journal |
| Volume | 122 |
| Issue number | 12 |
| DOIs | |
| State | Published - Jun 20 2023 |
Funding
This research was funded by NSF grants MCB-1817684 and MCB-2202202 to Z.B. B.S. was supported by a G-RISE Ph.D. traineeship from the National Institutes of Health (grant no. T32GM136499 ). We would like to acknowledge the Imaging Facility of Advanced Science Research Center at the CUNY Graduate Center for instrument use, scientific and technical assistance, and to thank Dr. Tong Wang for helpful discussions of the negative stain EM results. Some of this work was performed at the Simons Electron Microscopy Center at the New York Structural Biology Center (NYSBC), with major support from the Simons Foundation ( SF349247 ). We thank the staff members at the NYSBC for training the graduate student. Use of the Stanford Synchrotron Radiation Lightsource (SSRL), SLAC National Accelerator Laboratory, is supported by the U.S. Department of Energy , Office of Science , Office of Basic Energy Sciences under contract no. DE-AC02-76SF00515 . The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research , and by the National Institutes of Health, National Institute of General Medical Sciences ( P30GM133894 ). The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH. The Pilatus detector at Beamline 4-2 at SSRL was funded under National Institutes of Health grant S10OD021512 . This research used resources at the High Flux Isotope Reactor, a Department of Energy Office of Science User Facility operated by the Oak Ridge National Laboratory. This research was funded by NSF grants MCB-1817684 and MCB-2202202 to Z.B. B.S. was supported by a G-RISE Ph.D. traineeship from the National Institutes of Health (grant no. T32GM136499). We would like to acknowledge the Imaging Facility of Advanced Science Research Center at the CUNY Graduate Center for instrument use, scientific and technical assistance, and to thank Dr. Tong Wang for helpful discussions of the negative stain EM results. Some of this work was performed at the Simons Electron Microscopy Center at the New York Structural Biology Center (NYSBC), with major support from the Simons Foundation (SF349247). We thank the staff members at the NYSBC for training the graduate student. Use of the Stanford Synchrotron Radiation Lightsource (SSRL), SLAC National Accelerator Laboratory, is supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences under contract no. DE-AC02-76SF00515. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research, and by the National Institutes of Health, National Institute of General Medical Sciences (P30GM133894). The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH. The Pilatus detector at Beamline 4-2 at SSRL was funded under National Institutes of Health grant S10OD021512. This research used resources at the High Flux Isotope Reactor, a Department of Energy Office of Science User Facility operated by the Oak Ridge National Laboratory. The authors declare no competing interests.
| Funders | Funder number |
|---|---|
| Imaging Facility of Advanced Science Research Center | |
| NYSBC | |
| National Science Foundation | MCB-2202202, MCB-1817684 |
| National Institutes of Health | T32GM136499 |
| U.S. Department of Energy | |
| National Institute of General Medical Sciences | P30GM133894, S10OD021512 |
| Simons Foundation | SF349247 |
| Office of Science | |
| Basic Energy Sciences | DE-AC02-76SF00515 |
| Biological and Environmental Research | |
| Oak Ridge National Laboratory | |
| SLAC National Accelerator Laboratory |
Keywords
- adherens junction complex
- disordered protein
- small angle neutron scattering
- small angle x-ray scattering
- vinculin