Genetic analysis of the Complexin trans-clamping model for cross-linking SNARE complexes in vivo
DC Element | Wert | Sprache |
---|---|---|
dc.contributor.author | Cho, Richard W. | |
dc.contributor.author | Kuemmel, Daniel | |
dc.contributor.author | Li, Feng | |
dc.contributor.author | Baguley, Stephanie Wood | |
dc.contributor.author | Coleman, Jeff | |
dc.contributor.author | Rothman, James E. | |
dc.contributor.author | Littleton, J. Troy | |
dc.date.accessioned | 2021-12-23T16:18:39Z | - |
dc.date.available | 2021-12-23T16:18:39Z | - |
dc.date.issued | 2014 | |
dc.identifier.issn | 00278424 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/12787 | - |
dc.description.abstract | Complexin (Cpx) is a SNARE-binding protein that regulates neurotransmission by clamping spontaneous synaptic vesicle fusion in the absence of Ca2+ influx while promoting evoked release in response to an action potential. Previous studies indicated Cpx may cross-link multiple SNARE complexes via a trans interaction to function as a fusion clamp. During Ca2+ influx, Cpx is predicted to undergo a conformational switch and collapse onto a single SNARE complex in a cis-binding mode to activate vesicle release. To test this model in vivo, we performed structure-function studies of the Cpx protein in Drosophila. Using genetic rescue approaches with cpx mutants that disrupt SNARE cross-linking, we find that manipulations that are predicted to block formation of the trans SNARE array disrupt the clamping function of Cpx. Unexpectedly, these same mutants rescue action potential-triggered release, indicating trans-SNARE cross-linking by Cpx is not a prerequisite for triggering evoked fusion. In contrast, mutations that impair Cpx-mediated cis-SNARE interactions that are necessary for transition from an open to closed conformation fail to rescue evoked release defects in cpx mutants, although they clamp spontaneous release normally. Our in vivo genetic manipulations support several predictions made by the Cpx cross-linking model, but unexpected results suggest additional mechanisms are likely to exist that regulate Cpx's effects on SNARE-mediated fusion. Our findings also indicate that the inhibitory and activating functions of Cpx are genetically separable, and can be mapped to distinct molecular mechanisms that differentially regulate the SNARE fusion machinery. | |
dc.description.sponsorship | National Institutes of HealthUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USA [NS064750, GM 071458, NS40296]; NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCESUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of General Medical Sciences (NIGMS) [R01GM071458] Funding Source: NIH RePORTER; NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKEUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of Neurological Disorders & Stroke (NINDS) [F32NS064750, R01NS040296] Funding Source: NIH RePORTER; We thank Dr. Shyam Krishnakumar for helpful discussion, input, and careful reading of the manuscript. This work was supported by National Institutes of Health Grants NS064750 (to R. W. C.), GM 071458 (to J.E.R.), and NS40296 (to J.T.L.). | |
dc.language.iso | en | |
dc.publisher | NATL ACAD SCIENCES | |
dc.relation.ispartof | PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | |
dc.subject | CA2+ SENSOR | |
dc.subject | DOCKED VESICLES | |
dc.subject | DYNAMICS | |
dc.subject | EXOCYTOSIS | |
dc.subject | INTERMEDIATE | |
dc.subject | MEMBRANE-FUSION | |
dc.subject | Multidisciplinary Sciences | |
dc.subject | neurotransmitter release | |
dc.subject | PROTEINS | |
dc.subject | Science & Technology - Other Topics | |
dc.subject | SPONTANEOUS NEUROTRANSMITTER RELEASE | |
dc.subject | synapse | |
dc.subject | SYNAPTIC-VESICLE | |
dc.subject | SYNAPTOTAGMIN | |
dc.title | Genetic analysis of the Complexin trans-clamping model for cross-linking SNARE complexes in vivo | |
dc.type | journal article | |
dc.identifier.doi | 10.1073/pnas.1409311111 | |
dc.identifier.isi | ISI:000338985700066 | |
dc.description.volume | 111 | |
dc.description.issue | 28 | |
dc.description.startpage | 10317 | |
dc.description.endpage | 10322 | |
dc.contributor.orcid | 0000-0001-5576-2887 | |
dc.publisher.place | 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA | |
dcterms.isPartOf.abbreviation | Proc. Natl. Acad. Sci. U. S. A. | |
dcterms.oaStatus | Green Published, Bronze | |
crisitem.author.dept | FB 05 - Biologie/Chemie | - |
crisitem.author.deptid | fb05 | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.netid | KuDa343 | - |
Seitenaufrufe
2
Letzte Woche
0
0
Letzter Monat
0
0
geprüft am 29.05.2024