Classical protein kinase C(s) regulates targeting of synaptotagmin IX to the endocytic recycling compartment

Haberman, Yael; Ziv, Idit; Gorzalczany, Yaara; Fukuda, Mitsunori; Sagi‐Eisenberg, Ronit

doi:10.1242/jcs.02276

Cited by 28 publications

(39 citation statements)

References 32 publications

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“…2C). These two forms of Syt V may represent different posttranslational modifications, such as phosphorylation, as previously observed with Syt IV and Syt IX (36,37). We next compared the recruitment kinetics of Syt V and LAMP1 to phagosomes containing serum opsonizedzymozan by confocal microscopy.…”

Section: Syt V Associates To Phagosomesmentioning

confidence: 82%

The Exocytosis Regulator Synaptotagmin V Controls Phagocytosis in Macrophages

Vinet

Fukuda

Descoteaux

2008

The Journal of Immunology

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Synaptotagmins (Syts) play a key role in the regulation of Ca2+-triggered exocytosis and membrane fusion events, two crucial events associated to the phagocytic process. In the present study, we investigated the role of Syt V, a regulator of focal exocytosis, in phagocytosis. In macrophages, Syt V is localized on recycling endosomes and on filopodia-like structures and is recruited to the nascent phagosomes independently of the phagocytic receptor engaged. Silencing of Syt V by RNA interference revealed a role for this protein for phagocytosis, particularly under conditions of high membrane demand. In contrast, silencing of Syt V had no effect on the recruitment of the lysosomal marker LAMP1 to phagosomes, indicating that phagosome maturation is not regulated by Syt V. Collectively, these results illustrate the importance of Syt V in the regulation of an important innate function of macrophages. Furthermore, our results are consistent with the concept that focal exocytosis of endocytic organelles is a key event in phagocytosis and suggest that Syt V regulates this process.

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Section: Syt V Associates To Phagosomesmentioning

confidence: 82%

The Exocytosis Regulator Synaptotagmin V Controls Phagocytosis in Macrophages

Vinet

Fukuda

Descoteaux

2008

The Journal of Immunology

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“…Many studies have implicated PKC in the internalization of specific receptors, transporters, and channels (22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32). Some of these studies have focused on direct phosphorylation of these molecules by PKC.…”

Section: Discussionmentioning

confidence: 99%

Sustained Receptor Stimulation Leads to Sequestration of Recycling Endosomes in a Classical Protein Kinase C- and Phospholipase D-dependent Manner

Idkowiak‐Baldys¹,

Baldys

Raymond

et al. 2009

Journal of Biological Chemistry

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Considerable insight has been garnered on initial mechanisms of endocytosis of plasma membrane proteins and their subsequent trafficking through the endosomal compartment. It is also well established that ligand stimulation of many plasma membrane receptors leads to their internalization. However, stimulus-induced regulation of endosomal trafficking has not received much attention. In previous studies, we showed that sustained stimulation of protein kinase C (PKC) with phorbol esters led to sequestration of recycling endosomes in a juxtanuclear region. In this study, we investigated whether G-protein-coupled receptors that activate PKC exerted effects on endosomal trafficking. Stimulation of cells with serotonin (5-hydroxytryptamine (5-HT)) led to sequestration of the 5-HT receptor (5-HT 2A R) into a Rab11-positive juxtanuclear compartment. This sequestration coincided with translocation of PKC as shown by confocal microscopy. Mechanistically the observed sequestration of 5-HT 2A R was shown to require continuous PKC activity because it was inhibited by pretreatment with classical PKC inhibitor Gö6976 and could be reversed by posttreatment with this inhibitor. In addition, classical PKC autophosphorylation was necessary for receptor sequestration. Moreover inhibition of phospholipase D (PLD) activity and inhibition of PLD1 and PLD2 using dominant negative constructs also prevented this process. Functionally this sequestration did not affect receptor desensitization or resensitization as measured by intracellular calcium increase. However, the PKC-and PLD-dependent sequestration of receptors resulted in co-sequestration of other plasma membrane proteins and receptors as shown for epidermal growth factor receptor and protease activated receptor-1. This led to heterologous desensitization of those receptors and diverted their cellular fate by protecting them from agonist-induced degradation. Taken together, these results demonstrate a novel role for sustained receptor stimulation in regulation of intracellular trafficking, and this process requires sustained stimulation of PKC and PLD.

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“…Indeed, there are several examples in the literature showing PKC-dependent sequestration of molecules such as the dopamine transporter (14,15), dopamine receptor (42), parathyroid hormone receptor (43), norepinephrine transporter (16), human cationic amino acid transporter (17), and synaptotagmin IX (18)) to a compartment that resembles the pericentrion. Although those studies focused on specific PKC/target interaction, our results would suggest that the effects of PKC in those cases as well may be part of the more generalized effects on recycling endosomes described in this study.…”

Section: Discussionmentioning

confidence: 99%

“…Rotmann et al (17) found that treatment of cells with PMA resulted in internalization (possibly to the pericentrion) of the human cationic amino acid transporter hCAT-1. Furthermore, synaptotagmin IX was shown to be internalized to a compartment that resembles the pericentrion in a PKC␣-dependent manner (18). All these data indicate that active PKC␣ can indeed selectively regulate sequestration of certain molecules from the cell surface.…”

Section: Members Of the Protein Kinase C (Pkc)mentioning

confidence: 91%

Dynamic Sequestration of the Recycling Compartment by Classical Protein Kinase C

Idkowiak‐Baldys

Becker

Kitatani

et al. 2006

Journal of Biological Chemistry

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It has been previously shown that upon sustained stimulation (30 -60 min) with phorbol esters, protein kinase C (PKC) ␣ and ␤II become sequestered in a juxtanuclear region, the pericentrion. The activation of PKC also results in sequestration of transferrin, suggesting a role for PKC in regulating endocytosis and sequestration of recycling components. In this work we characterize the pericentrion as a PKC-dependent subset of the recycling compartment. We demonstrate that upon sustained stimulation of PKC, both protein (CD59, caveolin) and possibly also lipid (Bodipy-GM1) cargo become sequestered in a PKC-dependent manner. This sequestration displayed a strict temperature requirement and was inhibited below 32°C. Treatment of cells with phorbol myristate acetate for 60 min led to the formation of a distinct membrane structure. PKC sequestration and pericentrion formation were blocked by hypertonic sucrose as well as by potassium depletion (inhibitors of clathrin-dependent endocytosis) but not by nystatin or filipin, which inhibit clathrin-independent pathways. Interestingly, it was also observed that some molecules that internalize through clathrin-independent pathways (CD59, Bodipy-GM1, caveolin) also sequestered to the pericentrion upon sustained PKC activation, suggesting that PKC acted distal to the site of internalization of endocytic cargo. Together these results suggest that PKC regulates sequestration of recycling molecules into this compartment, the pericentrion.

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Classical protein kinase C(s) regulates targeting of synaptotagmin IX to the endocytic recycling compartment

Cited by 28 publications

References 32 publications

The Exocytosis Regulator Synaptotagmin V Controls Phagocytosis in Macrophages

The Exocytosis Regulator Synaptotagmin V Controls Phagocytosis in Macrophages

Sustained Receptor Stimulation Leads to Sequestration of Recycling Endosomes in a Classical Protein Kinase C- and Phospholipase D-dependent Manner

Dynamic Sequestration of the Recycling Compartment by Classical Protein Kinase C

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