The Slp Homology Domain of Synaptotagmin-like Proteins 1–4 and Slac2 Functions as a Novel Rab27A Binding Domain

Kuroda, Taruho S.; Fukuda, Mitsunori; Ariga, Hiroyoshi; Mikoshiba, Katsuhiko

doi:10.1074/jbc.m112414200

Cited by 206 publications

(275 citation statements)

References 46 publications

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“…Noticeably, a specific interaction between JFC1 and Rab27A has been demonstrated recently by Seabra and coworkers (13). Moreover, similar results have been described for a mouse homologous for JFC1 by a different group (30). Because it is well established that the Rab family of proteins function in membrane trafficking by determining the specificity of vesicle transport in eukaryotic cells (31), the findings described above confirm our previous suggestions.…”

Section: Discussionsupporting

confidence: 81%

The C2A domain of JFC1 binds to 3′-phosphorylated phosphoinositides and directs plasma membrane association in living cells

Catz

Johnson

Babior

2002

Proc. Natl. Acad. Sci. U.S.A.

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Phosphatidylinositol 3-kinase products play a central role in the regulation of several intracellular pathways via adaptor proteins that share the ability to bind to 3 -phosphoinositides with high affinity and specificity. JFC1 is a C2 domain-containing protein involved in cellular trafficking that has been shown to bind 3 -phosphoinositides in vitro. In this work, we demonstrate that the C2A domain of JFC1 is the module responsible for its binding to the plasma membrane via 3 -phosphoinositides in vivo. We show that the C2A domain of JFC1 is the only domain present in this protein that localizes to the plasma membrane in living cells. Moreover, the C2A domain of JFC1 binds 3 -phosphoinositides in vitro with similar specificity as that described for full-length JFC1, suggesting that the domain mediates the specific membrane localization of the full-length protein. Furthermore, the C2A domain of JFC1 colocalized with the pleckstrin homology domain of Akt in vivo, and both the JFC1 C2A domain and the full-length JFC1 dissociated from the membrane in the presence of PI 3-kinase specific inhibitors. We also show that the association of the C2A domain to the membrane is modulated by calcium. From these results we analyze possible mechanisms for the role of JFC1 in cellular trafficking. P hosphoinositides are specialized phospholipids known to localize to the inner leaflet of the plasma membrane, vesicles, and the nuclear envelope and participate in several signaling pathways (1). One of these phosphoinositides, phosphatidylinositol (4,5)-bisphosphate [PtdIns(4,5)P 2 ], plays a central role in the regulation of several intracellular pathways, from actin cytoskeleton modulation and vesicle trafficking (2) to modulation of the transcription factor Tubby protein (3). Furthermore, PtdIns(4,5)P 2 is a precursor for the second messenger phosphatidylinositol (3,4,5) trisphosphate [PtdIns(3,4,5)P 3 ] via the phosphoinositide 3-kinases ref. 4). PI 3-kinases and their products 3Ј-phosphorylated phosphoinositides (3Ј-phosphoinositides) are also involved in many crucial cell functions including vesicle trafficking (1, 2), cytoskeleton modulation, and cell survival (5). It is now clear that most of the functions regulated by phosphoinositides are mediated by adaptor proteins that share the ability to bind to PtdIns(4,5)P 2 and PI 3-kinase products with high affinity and specificity. These effectors are characterized by the presence of specialized phosphoinositide-binding domains. These domains include the pleckstrin homology (PH) domain (6), the FYVE finger (7), and the recently discovered PHOX homology (PX) domain (8). Furthermore, C2 domains have been shown to bind to phospholipids or phosphoinositides in the presence or absence of calcium (9). Proteins that contain C2 domains could generally be categorized as signal transduction mediators or membrane traffic effectors (10). The most extensively studied proteins included in the last group are the synaptotagmins (Syts). Syts are a group of transmembrane proteins that contain tan...

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Section: Discussionsupporting

confidence: 81%

The C2A domain of JFC1 binds to 3′-phosphorylated phosphoinositides and directs plasma membrane association in living cells

Catz

Johnson

Babior

2002

Proc. Natl. Acad. Sci. U.S.A.

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“…The antibody against insulin was from Linco Research (St. Charles, MO). The production of the antibodies against Slac2c/MyRIP and Myosin VIIa as well as the generation of the antibodies against Slp1, Slp2a, Slp3a, Slp5, and Slac2a/melanophilin have been described previously (ElAmraoui et al, 2002;Kuroda et al, 2002aKuroda et al, , 2002b. Fluorescently labeled secondary antibodies and Oregon green-coupled phalloidin were from Jackson ImmunoResearch Laboratories (West Grove, PA).…”

Section: Methodsmentioning

confidence: 99%

“…The mechanism of action of Rab27 in endocrine cells has been investigated only very recently and is still incompletely understood. Synaptotagmin-like proteins (Slp) and the related Slac2 proteins (Slps lacking C2 domains) are characterized by the presence of a unique amino terminal domain that confers the capacity to bind Rab27 (Kuroda et al, 2002a(Kuroda et al, , 2002b. One member of this protein family, Slp4/Granuphilin is localized on secretory granules of pancreatic ␤-cells and its overexpression causes a profound inhibition of insulin secretion (Wang et al, 1999;Coppola et al, 2002;.…”

Section: Introductionmentioning

confidence: 99%

Involvement of the Rab27 Binding Protein Slac2c/MyRIP in Insulin Exocytosis

Waselle

Coppola

Fukuda

et al. 2003

MBoC

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152

159

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“…Of the three subunits forming the tripartite complex, Mlph is by far the most versatile, because it has been shown to associate not only with Rab27a and MyoVa (20,21) but also with actin and the microtubule plus-end tracking EB1 protein via its C-terminal actin-binding domain (ABD) (22)(23)(24)(25). In vivo experiments initially argued for an important contribution of the ABD in the efficient distribution of melanosomes on the actin network (24).…”

mentioning

confidence: 99%

Myosin Va’s adaptor protein melanophilin enforces track selection on the microtubule and actin networks in vitro

Oberhofer

Spieler

Rosenfeld

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Pigment organelles, or melanosomes, are transported by kinesin, dynein, and myosin motors. As such, melanosome transport is an excellent model system to study the functional relationship between the microtubule-and actin-based transport systems. In mammalian melanocytes, it is well known that the Rab27a/melanophilin/myosin Va complex mediates actin-based transport in vivo. However, pathways that regulate the overall directionality of melanosomes on the actin/microtubule networks have not yet been delineated. Here, we investigated the role of PKA-dependent phosphorylation on the activity of the actin-based Rab27a/melanophilin/myosin Va transport complex in vitro. We found that melanophilin, specifically its C-terminal actin-binding domain (ABD), is a target of PKA. Notably, in vitro phosphorylation of the ABD closely recapitulated the previously described in vivo phosphorylation pattern. Unexpectedly, we found that phosphorylation of the ABD affected neither the interaction of the complex with actin nor its movement along actin tracks. Surprisingly, the phosphorylation state of melanophilin was instead important for reversible association with microtubules in vitro. Dephosphorylated melanophilin preferred binding to microtubules even in the presence of actin, whereas phosphorylated melanophilin associated with actin. Indeed, when actin and microtubules were present simultaneously, melanophilin's phosphorylation state enforced track selection of the Rab27a/melanophilin/ myosin Va transport complex. Collectively, our results unmasked the regulatory dominance of the melanophilin adaptor protein over its associated motor and offer an unexpected mechanism by which filaments of the cytoskeletal network compete for the moving organelles to accomplish directional transport on the cytoskeleton in vivo.melanophilin | myosin Va | intracellular transport | transport regulation

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The Slp Homology Domain of Synaptotagmin-like Proteins 1–4 and Slac2 Functions as a Novel Rab27A Binding Domain

Cited by 206 publications

References 46 publications

The C2A domain of JFC1 binds to 3′-phosphorylated phosphoinositides and directs plasma membrane association in living cells

The C2A domain of JFC1 binds to 3′-phosphorylated phosphoinositides and directs plasma membrane association in living cells

Involvement of the Rab27 Binding Protein Slac2c/MyRIP in Insulin Exocytosis

Myosin Va’s adaptor protein melanophilin enforces track selection on the microtubule and actin networks in vitro

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