Auto-regulation of Rab5 GEF activity in Rabex5 by allosteric structural changes, catalytic core dynamics and ubiquitin binding
Intracellular trafficking depends on the function of Rab GTPases, whose activation is regulated by guanine exchange factors (GEFs). The Rab5 GEF, Rabex5, was previously proposed to be auto-inhibited by its C-terminus. Here, we studied full-length Rabex5 and Rabaptin5 proteins as well as domain delet...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2019-11-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/46302 |
| _version_ | 1797998500159422464 |
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| author | Janelle Lauer Sandra Segeletz Alice Cezanne Giambattista Guaitoli Francesco Raimondi Marc Gentzel Vikram Alva Michael Habeck Yannis Kalaidzidis Marius Ueffing Andrei N Lupas Christian Johannes Gloeckner Marino Zerial |
| author_facet | Janelle Lauer Sandra Segeletz Alice Cezanne Giambattista Guaitoli Francesco Raimondi Marc Gentzel Vikram Alva Michael Habeck Yannis Kalaidzidis Marius Ueffing Andrei N Lupas Christian Johannes Gloeckner Marino Zerial |
| author_sort | Janelle Lauer |
| collection | DOAJ |
| description | Intracellular trafficking depends on the function of Rab GTPases, whose activation is regulated by guanine exchange factors (GEFs). The Rab5 GEF, Rabex5, was previously proposed to be auto-inhibited by its C-terminus. Here, we studied full-length Rabex5 and Rabaptin5 proteins as well as domain deletion Rabex5 mutants using hydrogen deuterium exchange mass spectrometry. We generated a structural model of Rabex5, using chemical cross-linking mass spectrometry and integrative modeling techniques. By correlating structural changes with nucleotide exchange activity for each construct, we uncovered new auto-regulatory roles for the ubiquitin binding domains and the Linker connecting those domains to the catalytic core of Rabex5. We further provide evidence that enhanced dynamics in the catalytic core are linked to catalysis. Our results suggest a more complex auto-regulation mechanism than previously thought and imply that ubiquitin binding serves not only to position Rabex5 but to also control its Rab5 GEF activity through allosteric structural alterations. |
| first_indexed | 2024-04-11T10:49:47Z |
| format | Article |
| id | doaj.art-59c0b315ce7c4837876fc46fb806c8fa |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T10:49:47Z |
| publishDate | 2019-11-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-59c0b315ce7c4837876fc46fb806c8fa2022-12-22T04:28:57ZengeLife Sciences Publications LtdeLife2050-084X2019-11-01810.7554/eLife.46302Auto-regulation of Rab5 GEF activity in Rabex5 by allosteric structural changes, catalytic core dynamics and ubiquitin bindingJanelle Lauer0https://orcid.org/0000-0003-1412-6766Sandra Segeletz1Alice Cezanne2Giambattista Guaitoli3Francesco Raimondi4Marc Gentzel5https://orcid.org/0000-0002-4482-6010Vikram Alva6https://orcid.org/0000-0003-1188-473XMichael Habeck7Yannis Kalaidzidis8Marius Ueffing9Andrei N Lupas10https://orcid.org/0000-0002-1959-4836Christian Johannes Gloeckner11https://orcid.org/0000-0001-6494-6944Marino Zerial12https://orcid.org/0000-0002-7490-4235Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, GermanyMax Planck Institute of Molecular Cell Biology and Genetics, Dresden, GermanyMax Planck Institute of Molecular Cell Biology and Genetics, Dresden, GermanyGerman Center for Neurodegenerative Diseases, Tübingen, GermanyBioquant, Heidelberg University, Heidelberg, Germany; Heidelberg University Biochemistry Centre (BZH), Heidelberg, GermanyMolecular Analysis-Mass Spectrometry Center for Molecular and Cellular Bioengineering, Technical University Dresden, Dresden, GermanyMax Planck Institute for Developmental Biology, Tuebingen, GermanyStatistical Inverse Problems in Biophysics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany; Felix Bernstein Institute for Mathematical Statistics in the Biosciences, University of Göttingen, Göttingen, GermanyMax Planck Institute of Molecular Cell Biology and Genetics, Dresden, GermanyCenter for Ophthalmology, Institute for Ophthalmic Research, University of Tübingen, Tübingen, GermanyMax Planck Institute for Developmental Biology, Tuebingen, GermanyGerman Center for Neurodegenerative Diseases, Tübingen, Germany; Center for Ophthalmology, Institute for Ophthalmic Research, University of Tübingen, Tübingen, GermanyMax Planck Institute of Molecular Cell Biology and Genetics, Dresden, GermanyIntracellular trafficking depends on the function of Rab GTPases, whose activation is regulated by guanine exchange factors (GEFs). The Rab5 GEF, Rabex5, was previously proposed to be auto-inhibited by its C-terminus. Here, we studied full-length Rabex5 and Rabaptin5 proteins as well as domain deletion Rabex5 mutants using hydrogen deuterium exchange mass spectrometry. We generated a structural model of Rabex5, using chemical cross-linking mass spectrometry and integrative modeling techniques. By correlating structural changes with nucleotide exchange activity for each construct, we uncovered new auto-regulatory roles for the ubiquitin binding domains and the Linker connecting those domains to the catalytic core of Rabex5. We further provide evidence that enhanced dynamics in the catalytic core are linked to catalysis. Our results suggest a more complex auto-regulation mechanism than previously thought and imply that ubiquitin binding serves not only to position Rabex5 but to also control its Rab5 GEF activity through allosteric structural alterations.https://elifesciences.org/articles/46302hydrogen deuterium exchange mass spectrometry (HDX-MS)guanine nucleotide exchange factor (GEF)Rab5 |
| spellingShingle | Janelle Lauer Sandra Segeletz Alice Cezanne Giambattista Guaitoli Francesco Raimondi Marc Gentzel Vikram Alva Michael Habeck Yannis Kalaidzidis Marius Ueffing Andrei N Lupas Christian Johannes Gloeckner Marino Zerial Auto-regulation of Rab5 GEF activity in Rabex5 by allosteric structural changes, catalytic core dynamics and ubiquitin binding eLife hydrogen deuterium exchange mass spectrometry (HDX-MS) guanine nucleotide exchange factor (GEF) Rab5 |
| title | Auto-regulation of Rab5 GEF activity in Rabex5 by allosteric structural changes, catalytic core dynamics and ubiquitin binding |
| title_full | Auto-regulation of Rab5 GEF activity in Rabex5 by allosteric structural changes, catalytic core dynamics and ubiquitin binding |
| title_fullStr | Auto-regulation of Rab5 GEF activity in Rabex5 by allosteric structural changes, catalytic core dynamics and ubiquitin binding |
| title_full_unstemmed | Auto-regulation of Rab5 GEF activity in Rabex5 by allosteric structural changes, catalytic core dynamics and ubiquitin binding |
| title_short | Auto-regulation of Rab5 GEF activity in Rabex5 by allosteric structural changes, catalytic core dynamics and ubiquitin binding |
| title_sort | auto regulation of rab5 gef activity in rabex5 by allosteric structural changes catalytic core dynamics and ubiquitin binding |
| topic | hydrogen deuterium exchange mass spectrometry (HDX-MS) guanine nucleotide exchange factor (GEF) Rab5 |
| url | https://elifesciences.org/articles/46302 |
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