A Complex Web of Signal-dependent Trafficking Underlies the Triorganellar Distribution of P-Selectin in Neuroendocrine PC12 Cells

Blagoveshchenskaya, Anastasiya D.; Hewitt, Eric W.; Cutler, Daniel F.

doi:10.1083/jcb.145.7.1419

Cited by 44 publications

(88 citation statements)

References 68 publications

(149 reference statements)

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“…In untreated cells, ZnT3-HA ( Figure 2B) or endogenous ZnT3 ( Figure 2F) were readily detectable in SLMV (fractions 12-15), yet BFA treatment substantially decreased the ZnT3 content in these fractions (Figure 2, C and G, n ϭ 3). In contrast, as described previously, the synaptophysin content was only minimally perturbed by BFA (Blagoveshchenskaya et al, 1999) (Figure 2, D and E), suggesting that different SLMV sorting mechanisms control synaptophysin and ZnT3. ZnT3 was redistributed from the membranes present in S1 to the P1 pellet.…”

Section: Znt3 Targeting To Pc12 Slmvcontrasting

confidence: 44%

“…Three criteria suggest that ZnT3 is targeted to SV by the AP-3 pathway: ZnT3 interacts with AP-3 through its carboxy-terminal tail in vitro, ZnT3 content in SV and larger brain membranes depends on the presence of functional AP-3, and ZnT3 subcellular distribution and targeting to SLMV are modified by brefeldin A. In contrast, synaptophysin targeting to SLMV is resistant to brefeldin A (Blagoveshchenskaya et al, 1999), and its sorting to brain SV remains unaltered in the absence of functional AP-3, thus suggesting that synaptophysin and ZnT3 preferentially follow distinct intracellular trafficking routes. However, in the absence of AP-3, a fifth of ZnT3 is still able to reach synaptic vesicles, implying that a proportion of ZnT3 can be sorted by an AP-3-independent mechanism.…”

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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

111

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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Section: Znt3 Targeting To Pc12 Slmvcontrasting

confidence: 44%

mentioning

confidence: 99%

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

111

178

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“…To determine the intracellular localization of this processing, we used a modified protocol for the isolation and separation of early/recycling endosomes from late endosomes in PC12 cells (8,64). Endogenous PS1 and mature nicastrin were found co-distributing with both markers of early/recycling (Rab5, transferrin receptor, and transferrin-HRP) and late endosomes (LAMP2, HRP) (Fig.…”

Section: Resultsmentioning

confidence: 99%

TrkA Receptor Activation by Nerve Growth Factor Induces Shedding of the p75 Neurotrophin Receptor Followed by Endosomal γ-Secretase-mediated Release of the p75 Intracellular Domain

Urra

Escudero²,

Ramos³

et al. 2007

Journal of Biological Chemistry

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Neurotrophins are trophic factors that regulate important neuronal functions. They bind two unrelated receptors, the Trk family of receptor-tyrosine kinases and the p75 neurotrophin receptor (p75). p75 was recently identified as a new substrate for ␥-secretase-mediated intramembrane proteolysis, generating a p75-derived intracellular domain (p75-ICD) with signaling capabilities. Using PC12 cells as a model, we studied how neurotrophins activate p75 processing and where these events occur in the cell. We demonstrate that activation of the TrkA receptor upon binding of nerve growth factor (NGF) regulates the metalloprotease-mediated shedding of p75 leaving a membranebound p75 C-terminal fragment (p75-CTF). Using subcellular fractionation to isolate a highly purified endosomal fraction, we demonstrate that p75-CTF ends up in endosomes where ␥-secretase-mediated p75-CTF cleavage occurs, resulting in the release of a p75-ICD. Moreover, we show similar structural requirements for ␥-secretase processing of p75 and amyloid precursor protein-derived CTFs. Thus, NGF-induced endocytosis regulates both signaling and proteolytic processing of p75.Neurotrophins belong to a small family of neurotrophic factors that include nerve growth factor (NGF), 4 brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT3), and neurotrophin-4 (NT4). They regulate different aspects of the developing and adult nervous system by binding to specific members of the Trk family of receptor-tyrosine kinases (TrkA, -B, and -C) or to the p75 neurotrophin receptor (p75). Their involvement includes neuronal migration, cell death, axonal elongation, myelinization, neuronal differentiation, and synaptic plasticity (1-3). p75 is a multifunctional type I transmembrane protein that is structurally related to the tumor necrosis factor receptor superfamily. It binds all neurotrophins, alone or in complex with Trk receptors, but also other ligands such as amyloid peptides and pro-neurotrophins (4, 5). The association of p75 with different receptors shapes the outcome of a signaling event. For example, binding of p75 to TrkA in the presence of NGF promotes neuronal survival, whereas interaction with the Nogo receptor (NgR) in the presence of its ligands, such as Nogo and myelin-associated glycoprotein (MAG), results in growth cone collapse and the inhibition of axonal regeneration (1, 4, 5). In addition, different downstream p75-associated signaling cascades are triggered through the interaction of its cytosolic portion with multiple adaptor proteins (6). Neurotrophin binding to p75 also induces accumulation of p75 in recycling endosomes where it associates with specific p75 downstream signaling effectors (7-9). Hence trafficking of p75 in the cell body or in axons (10, 11) may determine downstream signaling of p75 as previously shown for Trk receptors (12). The regulation of p75 signaling becomes even more complicated with the finding that p75 is subject to a dual processing starting with shedding of the ectodomain and followed by ␥-secretase cleavage. Thi...

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“…Although the retrieval of granule membranes to the Golgi complex has been extensively studied (23,24,31 -34) studies on sorting of individual granule membrane proteins at the level of the TGN are rare. However, we have recently been able to show that P-selectin is sorted into immature granules in PC12 cells by means of a tyrosine-based signal localised within its cytoplasmic domain (19). We have not yet determined whether this signal also operates in the same way in HUVECs, although since WPBlike structures can be seen budding from the TGN by electron microscopy (35), most likely P-selectin is being sorted within the TGN into the forming granule.…”

Section: Traffic 2000: 1: 783-793mentioning

confidence: 99%

Weibel‐Palade Body Membrane Proteins Exhibit Differential Trafficking After Exocytosis in Endothelial Cells

Arribas

Cutler

2000

Traffic

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Weibel-Palade bodies, the secretory granules of endothelial cells, possess two different membrane proteins. However, P-selectin is seen only in Weibel-Palade bodies in HUVECs, whereas CD63 is also seen in late endosomes/lysosomes. Since P-selectin is targeted to lysosomes in heterologous expression studies, we have determined whether a lysosomal targeting signal also operates within HUVECs. We have also examined the trafficking of CD63 to its two different intracellular locations. By following antibodies bound at the plasma membrane during stimulation, we have discovered that while half of the P-selectin recycles to the WPBs, 50% is rapidly delivered to a lamp-1-positive compartment. Thus, the lysosomal targeting signal of this protein also operates in HUVECs. CD63 is found constitutively at the cell surface of HUVECs and most of it is delivered to the late endosomes/lysosomes after internalisation. However, stimulation causes both a rise in the CD63 plasma membrane level and in the amount that recycles to the WPBs. Our data strongly suggest that the CD63 that originates in the WPB preferentially recycles to the granule rather than being delivered to the late endosome/ lysosome, and that there are, therefore, two separate pools of this protein within HUVECs. Our findings indicate that although P-selectin and CD63 are both targeted to the same compartments from the PM, the kinetics and the ratio of their targeting to Weibel-Palade bodies versus lysosomes are very different.

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A Complex Web of Signal-dependent Trafficking Underlies the Triorganellar Distribution of P-Selectin in Neuroendocrine PC12 Cells

Cited by 44 publications

References 68 publications

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

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

TrkA Receptor Activation by Nerve Growth Factor Induces Shedding of the p75 Neurotrophin Receptor Followed by Endosomal γ-Secretase-mediated Release of the p75 Intracellular Domain

Weibel‐Palade Body Membrane Proteins Exhibit Differential Trafficking After Exocytosis in Endothelial Cells

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