A v-SNARE participates in synaptic vesicle formation mediated by the AP3 adaptor complex

Salem, Natalie; Faúndez, Víctor; Horng, Jim-Tong; Kelly, Regis B.

doi:10.1038/2787

Cited by 86 publications

(69 citation statements)

References 37 publications

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“…38). Cleavage of vesicle-associated membrane protein caused defective synaptic vesicle biogenesis in PC12 cells (39). The SNARE protein also participates in COPII vesicle formation (40).…”

Section: Discussionmentioning

confidence: 99%

Reticulon 3 Binds the 2C Protein of Enterovirus 71 and Is Required for Viral Replication

Tang¹,

Yang²,

Wu³

et al. 2007

Journal of Biological Chemistry

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137

124

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Enterovirus 71 is an enterovirus of the family Picornaviridae. The 2C protein of poliovirus, a relative of enterovirus 71, is essential for viral replication. The poliovirus 2C protein is associated with host membrane vesicles, which form viral replication complexes where viral RNA synthesis takes place. We have now identified a host-encoded 2C binding protein called reticulon 3, which we found to be associated with the replication complex through direct interaction with the enterovirus 71-encoded 2C protein. We observed that the N terminus of the 2C protein, which has both RNA-and membrane-binding activity, interacted with reticulon 3. This region of interaction was mapped to its reticulon homology domain, whereas that of 2C was encoded by the 25th amino acid, isoleucine. Reticulon 3 could also interact with the 2C proteins encoded by other enteroviruses, such as poliovirus and coxsackievirus A16, implying that it is a common factor for such viral replication. Reduced production of reticulon 3 by RNA interference markedly reduced the synthesis of enterovirus 71-encoded viral proteins and replicative doublestranded RNA, reducing plaque formation and apoptosis. Furthermore, reintroduction of nondegradable reticulon 3 into these knockdown cells rescued enterovirus 71 infectivity, and viral protein and double-stranded RNA synthesis. Thus, reticulon 3 is an important component of enterovirus 71 replication, through its potential role in modulation of the sequential interactions between enterovirus 71 viral RNA and the replication complex.

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“…38). Cleavage of vesicle-associated membrane protein caused defective synaptic vesicle biogenesis in PC12 cells (39). The SNARE protein also participates in COPII vesicle formation (40).…”

Section: Discussionmentioning

confidence: 99%

Reticulon 3 Binds the 2C Protein of Enterovirus 71 and Is Required for Viral Replication

Tang¹,

Yang²,

Wu³

et al. 2007

Journal of Biological Chemistry

Self Cite

137

124

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“…VAMP II bearing the mutation N49A exhibits an enhanced AP-3-dependent sorting to SLMV compared with wild type, and thus should be enriched in vesicles derived from the AP-3 pathway (Grote et al, 1995;Salem et al, 1998). We magnetically isolated SLMV purified from PC12 VAMP II N49A cells by using well-characterized monoclonal antibodies against the cytosolic tails of synaptophysin (Wiedenmann and Franke, 1985) and VAMP II (Baumert et al, 1989).…”

Section: Znt3 and Synaptophysin Are Enriched In Different Slmv Subpopmentioning

confidence: 99%

“…The protein composition of the isolated vesicles was assessed by immunoblot and compared with the contents of the pooled purified SLMV. To detect differences in the distribution of SV antigens among vesicle subpopulations, reactions were designed so that the beads contained sufficient antibody to bind 20 -40% of the vesicle input ( Figure 5, A and B).VAMP II bearing the mutation N49A exhibits an enhanced AP-3-dependent sorting to SLMV compared with wild type, and thus should be enriched in vesicles derived from the AP-3 pathway (Grote et al, 1995;Salem et al, 1998). We magnetically isolated SLMV purified from PC12 VAMP II N49A cells by using well-characterized monoclonal antibodies against the cytosolic tails of synaptophysin (Wiedenmann and Franke, 1985) and VAMP II (Baumert et al, 1989).…”

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confidence: 99%

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The Zinc Transporter ZnT3 Interacts with AP-3 and It Is Preferentially Targeted to a Distinct Synaptic Vesicle Subpopulation

Salazar

Love

Werner

et al. 2004

MBoC

110

178

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Synaptic vesicles (SV) are generated by two different mechanisms, one AP-2 dependent and one AP-3 dependent. It has been uncertain, however, whether these mechanisms generate SV that differ in molecular composition. We explored this hypothesis by analyzing the targeting of ZnT3 and synaptophysin both to PC12 synaptic-like microvesicles (SLMV) as well as SV isolated from wild-type and AP-3-deficient mocha brains. ZnT3 cytosolic tail interacted selectively with AP-3 in cell-free assays. Accordingly, pharmacological disruption of either AP-2-or AP-3-dependent SLMV biogenesis preferentially reduced synaptophysin or ZnT3 targeting, respectively; suggesting that these antigens were concentrated in different vesicles. As predicted, immuno-isolated SLMV revealed that ZnT3 and synaptophysin were enriched in different vesicle populations. Likewise, morphological and biochemical analyses in hippocampal neurons indicated that these two antigens were also present in distinct but overlapping domains. ZnT3 SV content was reduced in AP-3-deficient neurons, but synaptophysin was not altered in the AP-3 null background. Our evidence indicates that neuroendocrine cells assemble molecularly heterogeneous SV and suggests that this diversity could contribute to the functional variety of synapses. INTRODUCTIONThe molecular diversity in total brain synaptic vesicle (SV) composition is generally presumed to result from differential expression of synaptic vesicle membrane proteins in different brain regions. However, the possibility that synaptic vesicles differ in composition because of different biogenesis mechanisms has not been explored. Different vesiculation pathways could result in molecularly diverse synaptic vesicles. Vesiculation mechanisms are known to produce distinct cargo carriers from a population of donor membranes (Bonifacino and Dell'Angelica, 1999;Springer et al., 1999;Boehm and Bonifacino, 2001). This process is achieved by adaptor complexes that recognize and concentrate specific membrane proteins in the donor membranes (Bonifacino and Dell 'Angelica, 1999;Kirchhausen, 1999). PC12 cells have been shown to possess two such adaptor-dependent pathways for the assembly of synaptic vesicles, also known as synaptic-like microvesicles (SLMV) (Shi et al., 1998;de Wit et al., 1999;Jarousse and Kelly, 2001). In one pathway, SLMV are generated from the plasma membrane by a vesiculation mechanism that requires the adaptor AP-2, clathrin, and the GTPase dynamin (Shi et al., 1998). In the second pathway, SLMV are generated from endosomes by the adaptor complex AP-3 (Faundez et al., 1998;Blumstein et al., 2001) and the GTPase ARF1 (Faundez et al., 1997). In contrast to the plasma membrane pathway, the endosomederived mechanism is highly sensitive to brefeldin A (BFA) (Shi et al., 1998).Phenotypes observed in the AP-3-deficient mocha mouse are consistent with a role for the AP-3-dependent, endosome-derived pathway in neurons (Kantheti et al., 1998). The mocha mossy fibers are devoid of both the synaptic vesiclespecific zinc transpor...

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“…Anti-116-kDa subunit of the vacuolar ATPase and VAMPII (69.1) were purchased from Synaptic Systems (Gö ttingen, Germany). AP-3 polyclonal antibodies, Arf1, and affinity-purified zinc transporter 3 (ZnT3) antibodies and sera have been already described (Salem et al, 1998;Faundez and Kelly, 2000;Salazar et al, 2004b). Monospecific affinity-purified polyclonal antibodies against phosphatidylinositol-4-kinase type II␣ were reported by Guo et al (2003).…”

Section: Antibodiesmentioning

confidence: 99%

Phosphatidylinositol-4-Kinase Type II α Is a Component of Adaptor Protein-3-derived Vesicles

Salazar

Craige

Wainer

et al. 2005

MBoC

100

136

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A membrane fraction enriched in vesicles containing the adaptor protein (AP) -3 cargo zinc transporter 3 was generated from PC12 cells and was used to identify new components of these organelles by mass spectrometry. Proteins prominently represented in the fraction included AP-3 subunits, synaptic vesicle proteins, and lysosomal proteins known to be sorted in an AP-3-dependent way or to interact genetically with AP-3. A protein enriched in this fraction was phosphatidylinositol-4-kinase type II␣ (PI4KII␣). Biochemical, pharmacological, and morphological analyses supported the presence of PI4KII␣ in AP-3-positive organelles. Furthermore, the subcellular localization of PI4KII␣ was altered in cells from AP-3-deficient mocha mutant mice. The PI4KII␣ normally present both in perinuclear and peripheral organelles was substantially decreased in the peripheral membranes of AP-3-deficient mocha fibroblasts. In addition, as is the case for other proteins sorted in an AP-3-dependent way, PI4KII␣ content was strongly reduced in nerve terminals of mocha hippocampal mossy fibers. The functional relationship between AP-3 and PI4KII␣ was further explored by PI4KII␣ knockdown experiments. Reduction of the cellular content of PI4KII␣ strongly decreased the punctate distribution of AP-3 observed in PC12 cells. These results indicate that PI4KII␣ is present on AP-3 organelles where it regulates AP-3 function. INTRODUCTIONMembrane-enclosed organelles possess distinctive protein compositions that are dynamically maintained by inbound and outbound vesicle carriers. These vesicles selectively concentrate appropriate membrane proteins while leaving behind resident proteins found in the donor organelle, a process called sorting (Bonifacino and Glick, 2004). Central to membrane protein sorting and vesiculation are a family of cytosolic coat complexes that mediate vesicle budding and function as cargo-specific adaptors. These include monomeric proteins and the heterotetrameric adaptor proteins (AP)-1, -2, -3, and -4 (Bonifacino and Glick, 2004;Robinson, 2004). These coats participate in the generation of vesicles that carry a unique array of membrane protein "cargoes." The characterization of these carriers has played a major role in the functional dissection of coat-dependent sorting and vesiculation mechanisms. For example, vesicles "in a basket" isolated from brain led to the biochemical identification of the first sorting machinery, clathrin and the AP-1 and AP-2 adaptors (Kanaseki and Kadota, 1969;Pearse, 1975;Pfeffer and Kelly, 1981;Pearse and Crowther, 1987). Subsequent studies of cargo molecules in brain clathrin-coated vesicles were crucial in revealing mechanisms of synaptic vesicle recycling (Pfeffer and Kelly, 1985;Maycox et al., 1992;Blondeau et al., 2004). The advent of complete genome sequencing and the concomitant development of informatics tools has led to the rapid identification of proteins by mass spectrometry (Taylor et al., 2003). Application of these methodologies to clathrin-coated vesicles has allowed the identifi...

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A v-SNARE participates in synaptic vesicle formation mediated by the AP3 adaptor complex

Cited by 86 publications

References 37 publications

Reticulon 3 Binds the 2C Protein of Enterovirus 71 and Is Required for Viral Replication

Reticulon 3 Binds the 2C Protein of Enterovirus 71 and Is Required for Viral Replication

The Zinc Transporter ZnT3 Interacts with AP-3 and It Is Preferentially Targeted to a Distinct Synaptic Vesicle Subpopulation

Phosphatidylinositol-4-Kinase Type II α Is a Component of Adaptor Protein-3-derived Vesicles

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