Abstract
The mechanochemical GTPase dynamin-related protein 1 (Drp1) catalyzes mitochondrial and peroxisomal fission, but the regulatory mechanisms remain ambiguous. Here we find that a conserved, intrinsically disordered, six-residue Short Linear Motif at the extreme Drp1 C-terminus, named CT-SLiM, constitutes a critical allosteric site that controls Drp1 structure and function in vitro and in vivo. Extension of the CT-SLiM by non-native residues, or its interaction with the protein partner GIPC-1, constrains Drp1 subunit conformational dynamics, alters self-assembly properties, and limits cooperative GTP hydrolysis, surprisingly leading to the fission of model membranes in vitro. In vivo, the involvement of the native CT-SLiM is critical for productive mitochondrial and peroxisomal fission, as both deletion and non-native extension of the CT-SLiM severely impair their progression. Thus, contrary to prevailing models, Drp1-catalyzed membrane fission relies on allosteric communication mediated by the CT-SLiM, deceleration of GTPase activity, and coupled changes in subunit architecture and assembly-disassembly dynamics.
| Original language | English |
|---|---|
| Article number | 52 |
| Number of pages | 17 |
| Journal | Nature Communications |
| Volume | 15 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2 Jan 2024 |
Bibliographical note
Funding Information:We thank Ashutosh Prince and Shane Wyborny (both of CWRU) for technical assistance in protein production. NIH R01 grants GM121583 and GM125844 supported work in the R.R. and J.A.M. laboratories, respectively. Work in the A.V.S. laboratory was supported by the PGC2018-099971-B-I00 and PID2021-127844NB-I00 grants funded by MCIN/AEI/10.13039/501100011033/ and by “ERDF A way of making Europe” and by the Basque Government Grant IT1625-22. Jon Agirre is a Royal Society University Research Fellow (award codes UF160039 and URF\R\221006). I.P.J. acknowledges the predoctoral fellowship from the University of the Basque Country. We are grateful for computational support from the University of York High Performance Computing service, Viking and the Research Computing team, notably Jasper Grimm and Emma Barnes. The authors are grateful to the Electron Microscopy and Crystallography platform of the CIC bioGUNE and the Basque Resource for Electron Microscopy (BREM) of the Biofisika Institute for providing access to cryo EM sample preparation and analysis equipment. Molecular graphics images were produced using the UCSF Chimera package from the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIH P41 RR-01081). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357. BioCAT was supported by grant P30 GM138395 from the National Institute of General Medical Sciences of the National Institutes of Health.
Funding Information:
We thank Ashutosh Prince and Shane Wyborny (both of CWRU) for technical assistance in protein production. NIH R01 grants GM121583 and GM125844 supported work in the R.R. and J.A.M. laboratories, respectively. Work in the A.V.S. laboratory was supported by the PGC2018-099971-B-I00 and PID2021-127844NB-I00 grants funded by MCIN/AEI/10.13039/501100011033/ and by “ERDF A way of making Europe” and by the Basque Government Grant IT1625-22. Jon Agirre is a Royal Society University Research Fellow (award codes UF160039 and URF\R\221006). I.P.J. acknowledges the predoctoral fellowship from the University of the Basque Country. We are grateful for computational support from the University of York High Performance Computing service, Viking and the Research Computing team, notably Jasper Grimm and Emma Barnes. The authors are grateful to the Electron Microscopy and Crystallography platform of the CIC bioGUNE and the Basque Resource for Electron Microscopy (BREM) of the Biofisika Institute for providing access to cryo EM sample preparation and analysis equipment. Molecular graphics images were produced using the UCSF Chimera package from the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIH P41 RR-01081). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357. BioCAT was supported by grant P30 GM138395 from the National Institute of General Medical Sciences of the National Institutes of Health.
Publisher Copyright:
© 2024, The Author(s).