A targeting signal in VAMP regulating transport to synaptic vesicles

Grote, Eric; Hao, Joe C.; Bennett, Mark K.; Kelly, Regis B.

doi:10.1016/0092-8674(95)90079-9

Cited by 158 publications

(234 citation statements)

References 34 publications

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“…Collectively these data suggest a key role for substrate residues Nterminal to the cleavage site as determinants of the specificity of BoNT/B. This region of VAMP has also been identified as a conserved signal sequence which plays a role in the regulation of VAMP transport to synaptic vesicles [18,19].…”

Section: Discussionmentioning

confidence: 86%

Substrate residues N‐terminal to the cleavage site of botulinum type B neurotoxin play a role in determining the specificity of its endopeptidase activity

Wictome¹,

Rossetto

Montecucco

et al. 1996

FEBS Letters

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Clostridium botulinum type B neurotoxin is a highly specific zinc-endopeptidase which cleaves vesicle-associated membrane protein (VAMP/synaptobrevin), a critical component of the vesicle docking/fusion mechanism. In this study, substrate residues flanking the N-terminal side of the cleavage site are shown to play a key role in enzyme substrate recognition. Two aspartate residues in this region are identified as critical determinants of the neurotoxin's specificity. These findings are discussed in relation to the mechanism by which botulinum type B neurotoxin cleaves its substrate.

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Section: Discussionmentioning

confidence: 86%

Substrate residues N‐terminal to the cleavage site of botulinum type B neurotoxin play a role in determining the specificity of its endopeptidase activity

Wictome¹,

Rossetto

Montecucco

et al. 1996

FEBS Letters

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“…In other studies, we have observed significant differences between the role of targeting signals in PC12 cells and in our in vivo assays. Specifically, we examined the role of previously defined SV targeting signals of synaptobrevin (Grote et al, 1995). We introduced into the C. elegans synaptobrevin gene two specific lesions (M46A and N49A), which altered targeting of synaptobrevin in PC12 cells.…”

Section: Caenorhabditis Elegans As a Model System To Study Protein Tamentioning

confidence: 99%

“…In keeping with this idea multiple signal sequences have been found within the cytoplasmic domain of synaptobrevin and synaptotagmin. In neuroendocrine PC12 cells, a sorting signal of synaptobrevin consisting of residues 41-50 probably functions at the plasma membrane because its deletion prevented endocytosis and SLMV targeting (Grote et al, 1995;Grote and Kelly, 1996). In contrast, residues 31-38 of synaptobrevin are required at a sorting stage other than endocytosis at the plasma membrane because a deletion mutant lacking these amino acids was endocytosed normally but excluded from SLMV (Grote et al, 1995).…”

Section: Synaptic Localization Signals In Other Systemsmentioning

confidence: 99%

“…In neuroendocrine PC12 cells, a sorting signal of synaptobrevin consisting of residues 41-50 probably functions at the plasma membrane because its deletion prevented endocytosis and SLMV targeting (Grote et al, 1995;Grote and Kelly, 1996). In contrast, residues 31-38 of synaptobrevin are required at a sorting stage other than endocytosis at the plasma membrane because a deletion mutant lacking these amino acids was endocytosed normally but excluded from SLMV (Grote et al, 1995). Although these regions are within the amphipathic helix required for biochemically defined synaptobrevin functions (SNARE complex formation), studies by Hao et al (1997) revealed little correlation between either endocytosis or SLMV targeting activity and complex formation activity.…”

Section: Synaptic Localization Signals In Other Systemsmentioning

confidence: 99%

“…Several studies have identified domains and motifs necessary for correct localization of SV proteins. Targeting and sorting signals have been identified in synaptophysin (Linstedt and Kelly, 1991), synaptobrevin (Grote et al, 1995;Grote and Kelly, 1996), synaptotagmin (Blagoveshchenskaya et al, 1999;Krasnov and Enikolopov, 2000), and vesicular neurotransmitter transporters (Tan et al, 1998;Varoqui and Erickson, 1998). However, no common motif that could serve as a universal targeting signal has been found by experimental studies or by direct comparison of the primary sequences of SV proteins.…”

Section: Introductionmentioning

confidence: 99%

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A Conserved Mechanism of Synaptogyrin Localization

Zhao

Nonet

2001

MBoC

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We have studied the localization of synaptogyrin family members in vivo. Both native and green fluorescent protein (GFP)-tagged Caenorhabditis elegans synaptogyrin (SNG-1) are expressed in neurons and synaptically localized. Deletion and mutational analysis with the use of GFP-tagged SNG-1 has defined a 38 amino acid sequence within the C terminus of SNG-1 and a single arginine in the cytoplasmic loop between transmembrane domain 2 and 3 that are required for SNG-1 localization. These domains may represent components of signals that target synaptogyrin for endocytosis from the plasma membrane and direct synaptogyrin to synaptic vesicles, respectively. In chimeric studies, these regions were sufficient to relocalize cellugyrin, a nonneuronal form of synaptogyrin, from nonsynaptic regions such as the sensory dendrites and the cell body to synaptic vesicles. Furthermore, GFP-tagged rat synaptogyrin is synaptically localized in neurons of C. elegans and in cultured hippocampal neurons. Similarly, the C-terminal domain of rat synaptogyrin is necessary for localization in hippocampal neurons. Our study suggests that the mechanisms for synaptogyrin localization are likely to be conserved from C. elegans to vertebrates. INTRODUCTIONSynaptic vesicles (SVs) contain a restricted set of membrane proteins important for neuronal function (Sü dhof, 1995;Calakos and Scheller, 1996;Fernandez-Chacon and Sü dhof, 1999). The mechanisms responsible for targeting these proteins specifically to the SV membrane are poorly understood. Integral membrane proteins are synthesized on the rough endoplasmic reticulum and traffic through the Golgi network. Proteins exiting the trans-Golgi network (TGN) are sorted to different types of transport vesicles. SV proteins are sorted to synaptic vesicle precursors (preSVs), which differ from mature SVs both in morphology and protein composition (Tsukita and Ishikawa, 1980; Okada et al., 1995). Evidence supports both direct routing of SV proteins from TGN to preSVs and indirect routing via the plasma membrane (reviewed by Hannah et al., 1999). preSVs are subsequently transported along axonal processes to the nerve terminal by motor proteins of the kinesin superfamily (reviewed by Hirokawa, 1998). How preSVs mature after delivery to the nerve terminal is unknown. preSVs might fuse with an endosomal compartment and SVs generated by budding from the endosome. A second, but not mutually exclusive possibility, is that preSVs are delivered to the presynaptic plasma membrane at the nerve terminal and then retrieved by the same mechanism(s) used to recycle SVs after regulated exocytosis (Hannah et al., 1999; reviewed by Buckley et al., 2000). Regardless of the exact route of trafficking to the mature organelle, SV proteins must be sorted away from other membrane proteins at several stages during their life cycle.Signal sequences resident in proteins are thought to mediate the sorting of many proteins to cellular compartments. Several studies have identified domains and motifs necessary for correct localizati...

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Selective localization and regulated release of calcitonin gene-related peptide from dense-core vesicles in engineered PC12 cells

1996

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Introduction of the gene for calcitonin into the neuroendocrine PC12 cell line resulted in the expression of the neuronal‐specific splice product, calcitonin gene‐related peptide (CGRP). Expression of this neuropeptide did not require treatment of the PC12 cells with NGF. By all available criteria, including biochemical, immunological, and morphological analysis, we have determined that the CGRP in stably transfected PC12 cells is sorted selectively into the large, dense‐core catecholamine‐containing secretory vesicles. Conversely, the CGRP is excluded from the small, synaptophysin‐rich vesicles present in the same cells. Stimulation conditions that trigger the release of catecholamines cause a parallel burst in the release of CGRP. In all these respects, the engineered PC12 cells process the foreign CGRP in a manner similar to that seen in spinal motor neurons in vivo. These results indicate that this small (37 amino acids) peptide contains sorting information sufficient for targeting to large, dense‐core vesicles in heterologous cells, placing very narrow constraints on the possible location of sorting signals. In addition, this CGRP‐expressing cell line opens the possibility of studying the physiological role of CGRP in the establishment and maintenance of neuromuscular contacts. © 1996 Wiley‐Liss, Inc.

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A targeting signal in VAMP regulating transport to synaptic vesicles

Cited by 158 publications

References 34 publications

Substrate residues N‐terminal to the cleavage site of botulinum type B neurotoxin play a role in determining the specificity of its endopeptidase activity

Substrate residues N‐terminal to the cleavage site of botulinum type B neurotoxin play a role in determining the specificity of its endopeptidase activity

A Conserved Mechanism of Synaptogyrin Localization

Selective localization and regulated release of calcitonin gene-related peptide from dense-core vesicles in engineered PC12 cells

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