UNC-18 and tomosyn antagonistically control synaptic vesicle priming downstream of UNC-13 in Caenorhabditis elegans

Seungmee Park, Na Ryum Bin, Bin Yu, Raymond Wong, Ewa Sitarska, Kyoko Sugita, Ke Ma, Junjie Xu, Chi Wei Tien, Arash Algouneh, Ekaterina Turlova, Siyan Wang, Pranay Siriya, Waleed Shahid, Lorraine Kalia, Zhong Ping Feng, Philippe P. Monnier, Hong Shuo Sun, Mei Zhen, Shangbang GaoJose Rizo-Rey, Shuzo Sugita

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Munc18-1/UNC-18 is believed to prime SNARE-mediated membrane fusion, yet the underlying mechanisms remain enigmatic. Here, we examine how potential gain-of-function mutations of Munc18-1/UNC-18 affect locomotory behavior and synaptic transmission, and how Munc18-1-mediated priming is related to Munc13-1/UNC-13 and Tomosyn/TOM-1, positive and negative SNARE regulators, respectively. We show that a Munc18-1(P335A)/UNC-18(P334A) mutation leads to significantly increased locomotory activity and acetylcholine release in Caenorhabditis elegans, as well as enhanced synaptic neurotransmission in cultured mammalian neurons. Importantly, similar to tom-1 null mutants, unc-18(P334A) mutants partially bypass the requirement of UNC-13. Moreover, unc-18(P334A) and tom-1 null mutations confer a strong synergy in suppressing the phenotypes of unc-13 mutants. Through biochemical experiments, we demonstrate that Munc18-1(P335A) exhibits enhanced activity in SNARE complex formation as well as in binding to the preformed SNARE complex, and partially bypasses the Munc13-1 requirement in liposome fusion assays. Our results indicate that Munc18-1/UNC-18 primes vesicle fusion downstream of Munc13-1/ UNC-13 by templating SNARE complex assembly and acts antagonistically with Tomosyn/TOM-1.

Original languageEnglish (US)
Pages (from-to)8797-8815
Number of pages19
JournalJournal of Neuroscience
Volume37
Issue number36
DOIs
StatePublished - Sep 6 2017

Fingerprint

SNARE Proteins
Synaptic Vesicles
Caenorhabditis elegans
Synaptic Transmission
Mutation
Membrane Fusion
Liposomes
Acetylcholine
Phenotype
Neurons

Keywords

  • C.elegans
  • Exocytosis
  • Membrane fusion
  • Munc18
  • SNARE
  • Synapse

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

UNC-18 and tomosyn antagonistically control synaptic vesicle priming downstream of UNC-13 in Caenorhabditis elegans. / Park, Seungmee; Bin, Na Ryum; Yu, Bin; Wong, Raymond; Sitarska, Ewa; Sugita, Kyoko; Ma, Ke; Xu, Junjie; Tien, Chi Wei; Algouneh, Arash; Turlova, Ekaterina; Wang, Siyan; Siriya, Pranay; Shahid, Waleed; Kalia, Lorraine; Feng, Zhong Ping; Monnier, Philippe P.; Sun, Hong Shuo; Zhen, Mei; Gao, Shangbang; Rizo-Rey, Jose; Sugita, Shuzo.

In: Journal of Neuroscience, Vol. 37, No. 36, 06.09.2017, p. 8797-8815.

Research output: Contribution to journalArticle

Park, S, Bin, NR, Yu, B, Wong, R, Sitarska, E, Sugita, K, Ma, K, Xu, J, Tien, CW, Algouneh, A, Turlova, E, Wang, S, Siriya, P, Shahid, W, Kalia, L, Feng, ZP, Monnier, PP, Sun, HS, Zhen, M, Gao, S, Rizo-Rey, J & Sugita, S 2017, 'UNC-18 and tomosyn antagonistically control synaptic vesicle priming downstream of UNC-13 in Caenorhabditis elegans', Journal of Neuroscience, vol. 37, no. 36, pp. 8797-8815. https://doi.org/10.1523/JNEUROSCI.0338-17.2017
Park, Seungmee ; Bin, Na Ryum ; Yu, Bin ; Wong, Raymond ; Sitarska, Ewa ; Sugita, Kyoko ; Ma, Ke ; Xu, Junjie ; Tien, Chi Wei ; Algouneh, Arash ; Turlova, Ekaterina ; Wang, Siyan ; Siriya, Pranay ; Shahid, Waleed ; Kalia, Lorraine ; Feng, Zhong Ping ; Monnier, Philippe P. ; Sun, Hong Shuo ; Zhen, Mei ; Gao, Shangbang ; Rizo-Rey, Jose ; Sugita, Shuzo. / UNC-18 and tomosyn antagonistically control synaptic vesicle priming downstream of UNC-13 in Caenorhabditis elegans. In: Journal of Neuroscience. 2017 ; Vol. 37, No. 36. pp. 8797-8815.
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abstract = "Munc18-1/UNC-18 is believed to prime SNARE-mediated membrane fusion, yet the underlying mechanisms remain enigmatic. Here, we examine how potential gain-of-function mutations of Munc18-1/UNC-18 affect locomotory behavior and synaptic transmission, and how Munc18-1-mediated priming is related to Munc13-1/UNC-13 and Tomosyn/TOM-1, positive and negative SNARE regulators, respectively. We show that a Munc18-1(P335A)/UNC-18(P334A) mutation leads to significantly increased locomotory activity and acetylcholine release in Caenorhabditis elegans, as well as enhanced synaptic neurotransmission in cultured mammalian neurons. Importantly, similar to tom-1 null mutants, unc-18(P334A) mutants partially bypass the requirement of UNC-13. Moreover, unc-18(P334A) and tom-1 null mutations confer a strong synergy in suppressing the phenotypes of unc-13 mutants. Through biochemical experiments, we demonstrate that Munc18-1(P335A) exhibits enhanced activity in SNARE complex formation as well as in binding to the preformed SNARE complex, and partially bypasses the Munc13-1 requirement in liposome fusion assays. Our results indicate that Munc18-1/UNC-18 primes vesicle fusion downstream of Munc13-1/ UNC-13 by templating SNARE complex assembly and acts antagonistically with Tomosyn/TOM-1.",
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T1 - UNC-18 and tomosyn antagonistically control synaptic vesicle priming downstream of UNC-13 in Caenorhabditis elegans

AU - Park, Seungmee

AU - Bin, Na Ryum

AU - Yu, Bin

AU - Wong, Raymond

AU - Sitarska, Ewa

AU - Sugita, Kyoko

AU - Ma, Ke

AU - Xu, Junjie

AU - Tien, Chi Wei

AU - Algouneh, Arash

AU - Turlova, Ekaterina

AU - Wang, Siyan

AU - Siriya, Pranay

AU - Shahid, Waleed

AU - Kalia, Lorraine

AU - Feng, Zhong Ping

AU - Monnier, Philippe P.

AU - Sun, Hong Shuo

AU - Zhen, Mei

AU - Gao, Shangbang

AU - Rizo-Rey, Jose

AU - Sugita, Shuzo

PY - 2017/9/6

Y1 - 2017/9/6

N2 - Munc18-1/UNC-18 is believed to prime SNARE-mediated membrane fusion, yet the underlying mechanisms remain enigmatic. Here, we examine how potential gain-of-function mutations of Munc18-1/UNC-18 affect locomotory behavior and synaptic transmission, and how Munc18-1-mediated priming is related to Munc13-1/UNC-13 and Tomosyn/TOM-1, positive and negative SNARE regulators, respectively. We show that a Munc18-1(P335A)/UNC-18(P334A) mutation leads to significantly increased locomotory activity and acetylcholine release in Caenorhabditis elegans, as well as enhanced synaptic neurotransmission in cultured mammalian neurons. Importantly, similar to tom-1 null mutants, unc-18(P334A) mutants partially bypass the requirement of UNC-13. Moreover, unc-18(P334A) and tom-1 null mutations confer a strong synergy in suppressing the phenotypes of unc-13 mutants. Through biochemical experiments, we demonstrate that Munc18-1(P335A) exhibits enhanced activity in SNARE complex formation as well as in binding to the preformed SNARE complex, and partially bypasses the Munc13-1 requirement in liposome fusion assays. Our results indicate that Munc18-1/UNC-18 primes vesicle fusion downstream of Munc13-1/ UNC-13 by templating SNARE complex assembly and acts antagonistically with Tomosyn/TOM-1.

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KW - C.elegans

KW - Exocytosis

KW - Membrane fusion

KW - Munc18

KW - SNARE

KW - Synapse

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