The SNARE Ykt6 mediates protein palmitoylation during an early stage of homotypic vacuole fusion

Dietrich, Lars E. P.; Gurezka, Rolf; Veit, Michael; Ungermann, Christian

doi:10.1038/sj.emboj.7600015

Cited by 72 publications

(89 citation statements)

References 69 publications

(139 reference statements)

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“…Antibodies to Ykt6 or the N-terminal longin domain interfered with palmitoylation at an early stage of fusion. Because in vitro palmitoylation required equimolar amounts of Ykt6 to modify Vac8, we suggested that acylation might occur by a nonenzymatic transfer mechanism (3,19). Such a mechanism has been suggested before and would explain previous autoacylation data (31).…”

Section: Discussionmentioning

confidence: 56%

“…Previous studies identified the SNARE Ykt6 as a factor that promotes Vac8 palmitoylation in vitro (19). Antibodies to Ykt6 or the N-terminal longin domain interfered with palmitoylation at an early stage of fusion.…”

Section: Discussionmentioning

confidence: 89%

“…3C). Previously, we reported that Vac8 palmitoylation occurs in an early subreaction on vacuoles that is sensitive to antibodies to Sec18 and Ykt6 (14,19). We therefore incubated vacuoles with [ 3 H]Pal-CoA and analyzed Vac8 palmitoylation.…”

Section: Pfa3 Affects Vacuole Fusion and Vac8mentioning

confidence: 99%

“…We have previously shown that the SNARE Ykt6 is involved in palmitoylation of vacuole-associated Vac8. Antibodies to Ykt6 block Vac8 palmitoylation, and purified Ykt6 can support in vitro palmitoylation of Vac8 when present in stoichiometric amounts (19). To search for additional factors involved in palmitoylation of vacuolar proteins, we determined whether any of the seven DHHC proteins are required for the localization and palmitoylation of Vac8, Yck3, or Meh1͞Ego1.…”

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confidence: 99%

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The DHHC protein Pfa3 affects vacuole-associated palmitoylation of the fusion factor Vac8

Hou

Subramanian

LaGrassa

et al. 2005

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

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Vacuole biogenesis depends on specific targeting and retention of peripheral membrane proteins. At least three palmitoylated proteins are found exclusively on yeast vacuoles: the fusion factor Vac8, the kinase Yck3, and a novel adaptor protein implicated in microautophagy, Meh1. Here, we analyze the role that putative acyltransferases of the DHHC family play in their localization and function. We find that Pfa3͞Ynl326c is required for efficient localization of Vac8 to vacuoles in vivo, while Yck3 or Meh1 localization is not impaired in any of the seven DHHC deletions. Vacuoleassociated Vac8 appears to be palmitoylated in a pfa3 mutant, but this population is refractive to further palmitoylation on isolated vacuoles. Vacuole morphology and inheritance, which both depend on Vac8 palmitoylation, appear normal, although there is a reduction in vacuole fusion. Interestingly, Pfa3 is required for the vacuolar localization of not only an SH4 domain that is targeted by myristate͞palmitate (as in Vac8) but also one that is targeted by a myristate͞basic stretch (as in Src). Our data indicate that Pfa3 has an important but not exclusive function for Vac8 localization to the vacuole.Yck3 ͉ SH4 domain ͉ acylation ͉ membrane targeting P rotein and lipid trafficking along the endomembrane system occurs by vesicular transport (1). Of all proteins implicated in vesicle fusion and fission, only a subset is permanently associated with membranes via transmembrane segments, whereas most are recruited to the membrane from the cytoplasm. The latter proteins depend on membrane receptors, lipids, or lipid anchors for binding to their appropriate target membrane (2). This poses the question of how the recruitment of these proteins is coordinated with their function.Palmitoylation has been discussed as a special lipid modification. It may direct proteins to specific membrane domains (3-5), and it is the only common lipid modification that is reversible, permitting cycling of a protein between membranes and cytosol. The identification and characterization of the underlying acylation͞deacylation machinery is therefore critical for understanding the function of palmitoylation. Recently, biochemical and genetic analyses have identified several proteins that are required for palmitoylation, including those of the so-called DHHC-CRD family of polytopic membrane proteins (6). Yeast contains seven homologs (Erf2, Swf1, Yol003c, Akr1, Akr2, Ydr459c, and Ynl326c͞Pfa3), which seem to be distributed throughout the endomembrane system as suggested by the GFP database and recent studies. At the ER, Swf1 is required for palmitoylation of Tlg1 (7), and Erf2 promotes Ras2 palmitoylation (8, 9). The Golgi-localized Akr1 is responsible for palmitoylation of the casein kinase I (CKI) isoform Yck2 (10). Similarly, several of the 20 or so mammalian DHHC proteins localize to distinct organelles, with critical roles for the palmitoylation of PSD-95, Ras, and SNAP-25 (11-13). This wide distribution of proteins involved in palmitoylation suggests that palmitoylati...

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Section: Discussionmentioning

confidence: 56%

Section: Discussionmentioning

confidence: 89%

Section: Pfa3 Affects Vacuole Fusion and Vac8mentioning

confidence: 99%

mentioning

confidence: 99%

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The DHHC protein Pfa3 affects vacuole-associated palmitoylation of the fusion factor Vac8

Hou

Subramanian

LaGrassa

et al. 2005

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

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“…In fact, the t-SNAREs syntaxins share a N-terminal H A H B H C domain necessary for viability and capable of intermolecular inhibition by binding to the Q-SNARE CCD (Nicholson et al, 1998;Parlati et al, 1999;Munson et al, 2000). Moreover, the v-SNAREs longins are characterized by the LD with profilin-like structure (Gonzalez et al, 2001;Tochio et al, 2001); this domain is capable to regulate membrane fusion (Martinez-Arca et al, 2000 and2001), as subcellular targeting (Hasegawa et al, 2003) by interacting with the clathrin adaptor AP-3 in the case of TI-VAMP/VAMP7 (Martinez-Arca et al, 2003), and protein palmitoylation in the case of Ykt6p (Dietrich et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

VAMP subfamilies identified by specific R-SNARE motifs

Rossi

Picco

Vacca

et al. 2004

Biology of the Cell

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In eukaryotes, interactions among the a-helical coiled-coil domains (CCDs) of soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) play a pivotal role in mediating the fusion among vesicles and target membranes. Surface residues of such CCDs are major candidates to regulate the specificity of membrane fusion, as they may alter local charge at the interaction layers and surface of the fusion complex, possibly modulating its formation and/or the binding of non-SNARE regulatory factors. Based on alternate patterns in surface residues, we have identified two motifs which group vesicular SNAREs in two novel subfamilies: RG-SNAREs and RD-SNAREs. The RG-SNARE CCD is common to all members of the widely conserved family of long VAMPs or longins and to yeast and non-neuronal VAMPs, possibly mediating ″basic″ fusion mechanisms; instead, only synaptobrevins from Bilateria share an RD-SNARE CCD, which is likely to mediate interactions to specific, yet unknown, regulatory factors and/or be the landmark of rapid fusion reactions like that mediating the release of neurotransmitters.

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Intracellular Membrane Fusion

Xu¹,

Hay

2009

Trafficking Inside Cells

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The SNARE Ykt6 mediates protein palmitoylation during an early stage of homotypic vacuole fusion

Cited by 72 publications

References 69 publications

The DHHC protein Pfa3 affects vacuole-associated palmitoylation of the fusion factor Vac8

The DHHC protein Pfa3 affects vacuole-associated palmitoylation of the fusion factor Vac8

VAMP subfamilies identified by specific R-SNARE motifs

Intracellular Membrane Fusion

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