Cargando…
SH3 Domains Differentially Stimulate Distinct Dynamin I Assembly Modes and G Domain Activity
Dynamin I is a highly regulated GTPase enzyme enriched in nerve terminals which mediates vesicle fission during synaptic vesicle endocytosis. One regulatory mechanism involves its interactions with proteins containing Src homology 3 (SH3) domains. At least 30 SH3 domain-containing proteins bind dyna...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4687643/ https://www.ncbi.nlm.nih.gov/pubmed/26659814 http://dx.doi.org/10.1371/journal.pone.0144609 |
_version_ | 1782406652784279552 |
---|---|
author | Krishnan, Sai Collett, Michael Robinson, Phillip J. |
author_facet | Krishnan, Sai Collett, Michael Robinson, Phillip J. |
author_sort | Krishnan, Sai |
collection | PubMed |
description | Dynamin I is a highly regulated GTPase enzyme enriched in nerve terminals which mediates vesicle fission during synaptic vesicle endocytosis. One regulatory mechanism involves its interactions with proteins containing Src homology 3 (SH3) domains. At least 30 SH3 domain-containing proteins bind dynamin at its proline-rich domain (PRD). Those that stimulate dynamin activity act by promoting its oligomerisation. We undertook a systematic parallel screening of 13 glutathione-S-transferase (GST)-tagged endocytosis-related SH3 domains on dynamin binding, GTPase activity and oligomerisation. No correlation was found between dynamin binding and their potency to stimulate GTPase activity. There was limited correlation between the extent of their ability to stimulate dynamin activity and the level of oligomerisation, indicating an as yet uncharacterised allosteric coupling of the PRD and G domain. We examined the two variants, dynamin Iab and Ibb, which differ in the alternately splice middle domain α2 helix. They responded differently to the panel of SH3s, with the extent of stimulation between the splice variants varying greatly between the SH3s. This study reveals that SH3 binding can act as a heterotropic allosteric regulator of the G domain via the middle domain α2 helix, suggesting an involvement of this helix in communicating the PRD-mediated allostery. This indicates that SH3 binding both stabilises multiple conformations of the tetrameric building block of dynamin, and promotes assembly of dynamin-SH3 complexes with distinct rates of GTP hydrolysis. |
format | Online Article Text |
id | pubmed-4687643 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-46876432015-12-31 SH3 Domains Differentially Stimulate Distinct Dynamin I Assembly Modes and G Domain Activity Krishnan, Sai Collett, Michael Robinson, Phillip J. PLoS One Research Article Dynamin I is a highly regulated GTPase enzyme enriched in nerve terminals which mediates vesicle fission during synaptic vesicle endocytosis. One regulatory mechanism involves its interactions with proteins containing Src homology 3 (SH3) domains. At least 30 SH3 domain-containing proteins bind dynamin at its proline-rich domain (PRD). Those that stimulate dynamin activity act by promoting its oligomerisation. We undertook a systematic parallel screening of 13 glutathione-S-transferase (GST)-tagged endocytosis-related SH3 domains on dynamin binding, GTPase activity and oligomerisation. No correlation was found between dynamin binding and their potency to stimulate GTPase activity. There was limited correlation between the extent of their ability to stimulate dynamin activity and the level of oligomerisation, indicating an as yet uncharacterised allosteric coupling of the PRD and G domain. We examined the two variants, dynamin Iab and Ibb, which differ in the alternately splice middle domain α2 helix. They responded differently to the panel of SH3s, with the extent of stimulation between the splice variants varying greatly between the SH3s. This study reveals that SH3 binding can act as a heterotropic allosteric regulator of the G domain via the middle domain α2 helix, suggesting an involvement of this helix in communicating the PRD-mediated allostery. This indicates that SH3 binding both stabilises multiple conformations of the tetrameric building block of dynamin, and promotes assembly of dynamin-SH3 complexes with distinct rates of GTP hydrolysis. Public Library of Science 2015-12-10 /pmc/articles/PMC4687643/ /pubmed/26659814 http://dx.doi.org/10.1371/journal.pone.0144609 Text en © 2015 Krishnan et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Krishnan, Sai Collett, Michael Robinson, Phillip J. SH3 Domains Differentially Stimulate Distinct Dynamin I Assembly Modes and G Domain Activity |
title | SH3 Domains Differentially Stimulate Distinct Dynamin I Assembly Modes and G Domain Activity |
title_full | SH3 Domains Differentially Stimulate Distinct Dynamin I Assembly Modes and G Domain Activity |
title_fullStr | SH3 Domains Differentially Stimulate Distinct Dynamin I Assembly Modes and G Domain Activity |
title_full_unstemmed | SH3 Domains Differentially Stimulate Distinct Dynamin I Assembly Modes and G Domain Activity |
title_short | SH3 Domains Differentially Stimulate Distinct Dynamin I Assembly Modes and G Domain Activity |
title_sort | sh3 domains differentially stimulate distinct dynamin i assembly modes and g domain activity |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4687643/ https://www.ncbi.nlm.nih.gov/pubmed/26659814 http://dx.doi.org/10.1371/journal.pone.0144609 |
work_keys_str_mv | AT krishnansai sh3domainsdifferentiallystimulatedistinctdynaminiassemblymodesandgdomainactivity AT collettmichael sh3domainsdifferentiallystimulatedistinctdynaminiassemblymodesandgdomainactivity AT robinsonphillipj sh3domainsdifferentiallystimulatedistinctdynaminiassemblymodesandgdomainactivity |