Molecular mechanism of DRP1 assembly studied in vitro by cryo-electron microscopy.
Mitochondria are dynamic organelles that continually adapt their morphology by fusion and fission events. An imbalance between fusion and fission has been linked to major neurodegenerative diseases, including Huntington's, Alzheimer's, and Parkinson's diseases. A member of the Dynamin...
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doaj-aa73bacff3b64d489b08eb67af5a64572020-11-25T01:52:52ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01126e017939710.1371/journal.pone.0179397Molecular mechanism of DRP1 assembly studied in vitro by cryo-electron microscopy.Kaustuv BasuDriss LajoieTristan Aumentado-ArmstrongJin ChenRoman I KoningBlaise BossyMihnea BostinaAttila SikElla Bossy-WetzelIsabelle RouillerMitochondria are dynamic organelles that continually adapt their morphology by fusion and fission events. An imbalance between fusion and fission has been linked to major neurodegenerative diseases, including Huntington's, Alzheimer's, and Parkinson's diseases. A member of the Dynamin superfamily, dynamin-related protein 1 (DRP1), a dynamin-related GTPase, is required for mitochondrial membrane fission. Self-assembly of DRP1 into oligomers in a GTP-dependent manner likely drives the division process. We show here that DRP1 self-assembles in two ways: i) in the presence of the non-hydrolysable GTP analog GMP-PNP into spiral-like structures of ~36 nm diameter; and ii) in the presence of GTP into rings composed of 13-18 monomers. The most abundant rings were composed of 16 monomers and had an outer and inner ring diameter of ~30 nm and ~20 nm, respectively. Three-dimensional analysis was performed with rings containing 16 monomers. The single-particle cryo-electron microscopy map of the 16 monomer DRP1 rings suggests a side-by-side assembly of the monomer with the membrane in a parallel fashion. The inner ring diameter of 20 nm is insufficient to allow four membranes to exist as separate entities. Furthermore, we observed that mitochondria were tubulated upon incubation with DRP1 protein in vitro. The tubes had a diameter of ~ 30nm and were decorated with protein densities. These findings suggest DRP1 tubulates mitochondria, and that additional steps may be required for final mitochondrial fission.http://europepmc.org/articles/PMC5478127?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kaustuv Basu Driss Lajoie Tristan Aumentado-Armstrong Jin Chen Roman I Koning Blaise Bossy Mihnea Bostina Attila Sik Ella Bossy-Wetzel Isabelle Rouiller |
spellingShingle |
Kaustuv Basu Driss Lajoie Tristan Aumentado-Armstrong Jin Chen Roman I Koning Blaise Bossy Mihnea Bostina Attila Sik Ella Bossy-Wetzel Isabelle Rouiller Molecular mechanism of DRP1 assembly studied in vitro by cryo-electron microscopy. PLoS ONE |
author_facet |
Kaustuv Basu Driss Lajoie Tristan Aumentado-Armstrong Jin Chen Roman I Koning Blaise Bossy Mihnea Bostina Attila Sik Ella Bossy-Wetzel Isabelle Rouiller |
author_sort |
Kaustuv Basu |
title |
Molecular mechanism of DRP1 assembly studied in vitro by cryo-electron microscopy. |
title_short |
Molecular mechanism of DRP1 assembly studied in vitro by cryo-electron microscopy. |
title_full |
Molecular mechanism of DRP1 assembly studied in vitro by cryo-electron microscopy. |
title_fullStr |
Molecular mechanism of DRP1 assembly studied in vitro by cryo-electron microscopy. |
title_full_unstemmed |
Molecular mechanism of DRP1 assembly studied in vitro by cryo-electron microscopy. |
title_sort |
molecular mechanism of drp1 assembly studied in vitro by cryo-electron microscopy. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2017-01-01 |
description |
Mitochondria are dynamic organelles that continually adapt their morphology by fusion and fission events. An imbalance between fusion and fission has been linked to major neurodegenerative diseases, including Huntington's, Alzheimer's, and Parkinson's diseases. A member of the Dynamin superfamily, dynamin-related protein 1 (DRP1), a dynamin-related GTPase, is required for mitochondrial membrane fission. Self-assembly of DRP1 into oligomers in a GTP-dependent manner likely drives the division process. We show here that DRP1 self-assembles in two ways: i) in the presence of the non-hydrolysable GTP analog GMP-PNP into spiral-like structures of ~36 nm diameter; and ii) in the presence of GTP into rings composed of 13-18 monomers. The most abundant rings were composed of 16 monomers and had an outer and inner ring diameter of ~30 nm and ~20 nm, respectively. Three-dimensional analysis was performed with rings containing 16 monomers. The single-particle cryo-electron microscopy map of the 16 monomer DRP1 rings suggests a side-by-side assembly of the monomer with the membrane in a parallel fashion. The inner ring diameter of 20 nm is insufficient to allow four membranes to exist as separate entities. Furthermore, we observed that mitochondria were tubulated upon incubation with DRP1 protein in vitro. The tubes had a diameter of ~ 30nm and were decorated with protein densities. These findings suggest DRP1 tubulates mitochondria, and that additional steps may be required for final mitochondrial fission. |
url |
http://europepmc.org/articles/PMC5478127?pdf=render |
work_keys_str_mv |
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