In vivo analysis of conserved C. elegans tomosyn domains.
Neurosecretion is critically dependent on the assembly of a macromolecular complex between the SNARE proteins syntaxin, SNAP-25 and synaptobrevin. Evidence indicates that the binding of tomosyn to syntaxin and SNAP-25 interferes with this assembly, thereby negatively regulating both synaptic transmi...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3195084?pdf=render |
| _version_ | 1818961096614084608 |
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| author | Anna O Burdina Susan M Klosterman Ludmila Shtessel Shawn Ahmed Janet E Richmond |
| author_facet | Anna O Burdina Susan M Klosterman Ludmila Shtessel Shawn Ahmed Janet E Richmond |
| author_sort | Anna O Burdina |
| collection | DOAJ |
| description | Neurosecretion is critically dependent on the assembly of a macromolecular complex between the SNARE proteins syntaxin, SNAP-25 and synaptobrevin. Evidence indicates that the binding of tomosyn to syntaxin and SNAP-25 interferes with this assembly, thereby negatively regulating both synaptic transmission and peptide release. Tomosyn has two conserved domains: an N-terminal encompassing multiple WD40 repeats predicted to form two β-propeller structures and a C-terminal SNARE-binding motif. To assess the function of each domain, we performed an in vivo analysis of the N- and C- terminal domains of C. elegans tomosyn (TOM-1) in a tom-1 mutant background. We verified that both truncated TOM-1 constructs were transcribed at levels comparable to rescuing full-length TOM-1, were of the predicted size, and localized to synapses. Unlike full-length TOM-1, expression of the N- or C-terminal domains alone was unable to restore inhibitory control of synaptic transmission in tom-1 mutants. Similarly, co-expression of both domains failed to restore TOM-1 function. In addition, neither the N- nor C-terminal domain inhibited release when expressed in a wild-type background. Based on these results, we conclude that the ability of tomosyn to regulate neurotransmitter release in vivo depends on the physical integrity of the protein, indicating that both N- and C-terminal domains are necessary but not sufficient for effective inhibition of release in vivo. |
| first_indexed | 2024-12-20T12:08:00Z |
| format | Article |
| id | doaj.art-95c23c279e6848ff95b01456f7398940 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-20T12:08:00Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-95c23c279e6848ff95b01456f73989402022-12-21T19:41:20ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01610e2618510.1371/journal.pone.0026185In vivo analysis of conserved C. elegans tomosyn domains.Anna O BurdinaSusan M KlostermanLudmila ShtesselShawn AhmedJanet E RichmondNeurosecretion is critically dependent on the assembly of a macromolecular complex between the SNARE proteins syntaxin, SNAP-25 and synaptobrevin. Evidence indicates that the binding of tomosyn to syntaxin and SNAP-25 interferes with this assembly, thereby negatively regulating both synaptic transmission and peptide release. Tomosyn has two conserved domains: an N-terminal encompassing multiple WD40 repeats predicted to form two β-propeller structures and a C-terminal SNARE-binding motif. To assess the function of each domain, we performed an in vivo analysis of the N- and C- terminal domains of C. elegans tomosyn (TOM-1) in a tom-1 mutant background. We verified that both truncated TOM-1 constructs were transcribed at levels comparable to rescuing full-length TOM-1, were of the predicted size, and localized to synapses. Unlike full-length TOM-1, expression of the N- or C-terminal domains alone was unable to restore inhibitory control of synaptic transmission in tom-1 mutants. Similarly, co-expression of both domains failed to restore TOM-1 function. In addition, neither the N- nor C-terminal domain inhibited release when expressed in a wild-type background. Based on these results, we conclude that the ability of tomosyn to regulate neurotransmitter release in vivo depends on the physical integrity of the protein, indicating that both N- and C-terminal domains are necessary but not sufficient for effective inhibition of release in vivo.http://europepmc.org/articles/PMC3195084?pdf=render |
| spellingShingle | Anna O Burdina Susan M Klosterman Ludmila Shtessel Shawn Ahmed Janet E Richmond In vivo analysis of conserved C. elegans tomosyn domains. PLoS ONE |
| title | In vivo analysis of conserved C. elegans tomosyn domains. |
| title_full | In vivo analysis of conserved C. elegans tomosyn domains. |
| title_fullStr | In vivo analysis of conserved C. elegans tomosyn domains. |
| title_full_unstemmed | In vivo analysis of conserved C. elegans tomosyn domains. |
| title_short | In vivo analysis of conserved C. elegans tomosyn domains. |
| title_sort | in vivo analysis of conserved c elegans tomosyn domains |
| url | http://europepmc.org/articles/PMC3195084?pdf=render |
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