Vacuolar protein sorting receptor in Schizosaccharomyces pombe

Iwaki, Toru; Hosomi, Akíra; Tokudomi, Sanae; Kusunoki, Yoko; Fujita, Yuka; Giga-Hama, Yuko; Tanaka, Naotaka; Takegawa, Kaoru

doi:10.1099/mic.0.28627-0

Cited by 44 publications

(79 citation statements)

References 62 publications

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“…In Sch. pombe, it is also known that PX domain-containing proteins Vps5p and Vps17p are involved in vacuolar protein transport (Koga et al, 2004;Iwaki et al, 2006). Taken together, these facts seem to indicate that Vsl1p corresponds to Sac.…”

Section: Discussionsupporting

confidence: 53%

Vsl1p cooperates with Fsv1p for vacuolar protein transport and homotypic fusion in Schizosaccharomyces pombe

et al. 2015

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Members of the SNARE protein family participate in the docking-fusion step of several intracellular vesicular transport events. Saccharomyces cerevisiae Vam7p was identified as a SNARE protein that acts in vacuolar protein transport and membrane fusion. However, in Schizosaccharomyces pombe, there have been no reports regarding the counterpart of Vam7p. Here, we found that, although the SPCC594.06c gene has low similarity to Vam7p, the product of SPCC594.06c has a PX domain and SNARE motif like Vam7p, and thus we designated the gene Sch. pombe vsl1 + (Vam7-like protein 1). The vsl1D cells showed no obvious defect in vacuolar protein transport. However, cells of the vsl1D mutant with a deletion of fsv1 + , which encodes another SNARE protein, displayed extreme defects in vacuolar protein transport and vacuolar morphology. Vsl1p was localized to the vacuolar membrane and prevacuolar compartment, and its PX domain was essential for proper localization. Expression of the fusion protein GFP-Vsl1p was able to suppress ZnCl 2 sensitivity and the vacuolar protein sorting defect in the fsv1D cells. Moreover, GFP-Vsl1p was mislocalized in a pep12D mutant and in cells overexpressing fsv1 + . Importantly, overexpression of Sac. cerevisiae VAM7 could suppress the sensitivity to ZnCl 2 of vsl1D cells and the vacuolar morphology defect of vsl1Dfsv1D cells in Sch. pombe. Taken together, these data suggest that Vsl1p and Fsv1p are required for vacuolar protein transport and membrane fusion, and they function cooperatively with Pep12p in the same membrane-trafficking step.

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Section: Discussionsupporting

confidence: 53%

Vsl1p cooperates with Fsv1p for vacuolar protein transport and homotypic fusion in Schizosaccharomyces pombe

et al. 2015

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“…S4B in the supplemental material). To decipher in which Golgi compartments sp-Erf2 localizes, we examined markers for two key Golgi compartments, Anp1 for the cis-Golgi compartment (46) and sp-Sec72 and Gms1 for the trans-Golgi compartment (22,46,47). Vesicles are received from the ER by the cis-Golgi compartment and can be delivered to the PM from the trans-Golgi compartment.…”

Section: Resultsmentioning

confidence: 99%

Regulation of Ras Localization and Cell Transformation by Evolutionarily Conserved Palmitoyltransferases

Young

Zheng

Wilkins

et al. 2014

Molecular and Cellular Biology

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e Ras can act on the plasma membrane (PM) to mediate extracellular signaling and tumorigenesis. To identify key components controlling Ras PM localization, we performed an unbiased screen to seek Schizosaccharomyces pombe mutants with reduced PM Ras. Five mutants were found with mutations affecting the same gene, S. pombe erf2 (sp-erf2), encoding sp-Erf2, a palmitoyltransferase, with various activities. sp-Erf2 localizes to the trans-Golgi compartment, a process which is mediated by its third transmembrane domain and the Erf4 cofactor. In fission yeast, the human ortholog zDHHC9 rescues the phenotypes of sp-erf2 null cells. In contrast, expressing zDHHC14, another sp-Erf2-like human protein, did not rescue Ras1 mislocalization in these cells. Importantly, ZDHHC9 is widely overexpressed in cancers. Overexpressing ZDHHC9 promotes, while repressing it diminishes, Ras PM localization and transformation of mammalian cells. These data strongly demonstrate that sp-Erf2/zDHHC9 palmitoylates Ras proteins in a highly selective manner in the trans-Golgi compartment to facilitate PM targeting via the transGolgi network, a role that is most certainly critical for Ras-driven tumorigenesis. Mammals have three RAS genes, HRAS, NRAS, and KRAS, which encode membrane-bound GTPases that control a wide range of signal transduction pathways critical for growth, differentiation, and cytoskeleton remodeling (1). The best-known Ras function is to mediate growth factor signaling that occurs at the plasma membrane (PM), and deregulation of this and other Ras functions can promote tumor formation (2, 3). While all Ras proteins are farnesylated to gain general membrane affinity, a second signal is needed to specifically localize to the PM. For HRas and NRas, this signal is provided by palmitoylation at cysteine residues immediately upstream of the C-terminal CAAX motif. This reaction can be catalyzed by protein acyltransferases (PATs) that contain a DHHC domain rich with aspartate, histidine, and cysteine residues. These DHHC-PATs are expected to localize in the Golgi complex to allow the delivery of palmitoylated Ras proteins to the PM through the trans-Golgi system (4) although this concept has not been thoroughly demonstrated in vivo.Humans have 24 DHHC-PATs, but it remains unresolved which of these control palmitoylation of Ras proteins in vivo. In contrast, the Ras pathways are considerably simpler in yeasts. Early studies from the budding yeast Saccharomyces cerevisiae, which has seven DHHC-PATs and two Ras proteins, suggest that only S. cerevisiae Erf2 (sc-Erf2) controls palmitoylation of its Ras proteins (5, 6). By sequence alignment to find putative orthologs, zDHHC9 was predicted to be a human Ras PAT (7). Although zDHHC9 can catalyze Ras palmitoylation in vitro, zDHHC9 cannot rescue sc-erf2 mutant phenotypes in budding yeast (8), and until now there has been no evidence that zDHHC9 or other human DHHC-PATs can control Ras localization in mammalian cells. Furthermore, while ectopically expressed zDHHC9 localizes to the Golgi compl...

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“…Correct localization of Vps10p to Golgi is maintained by retrograde transport from PVC to Golgi, and a sorting complex called the retromer complex consisting of five Vps proteins (Vps5p, Vps17p, Vps26p, Vps29p, and Vps35p) is also conserved in S. pombe. 12,15) To determine what caused this sorting defect in aah3Á cells, cellular localization of a Vps10p was observed. A plasmid, pTN197/vps10, was transformed into wild-type and aah3Á cells to express GFP-fused Vps10p under the control of the thiamine-repressible and attenuated nmt1 promoter.…”

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confidence: 99%

“…8) cpy1Á (h þ , ura4-D18, leu1-32, ade6-M216, his2, cpy1::ura4 þ ) 10) was used as a negative control. vps34Á (h þ , ura4-D18, leu1-32, ade6-M216, his2, vps34::ura4 þ ) 11) and vps26Á (h À , ura4-C190T, leu1-32, vps26::ura4 þ ) 12) were used as positive controls secreting strong and moderate-weak levels of CPY respectively. Standard rich medium (YES) was used as described previously.…”

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confidence: 99%

“…13) Colony blot assay was performed as described previously. 12,14) Briefly, yeast strains were precultured in YES medium and then replica-plated on a fresh YES plate in contact with a nitrocellulose membrane. The plates were then incubated at 30 C for 2 d. The nitrocellulose membranes were washed several times with water and then subjected to The membranes were subjected to immunoblotting with rabbit anti-CPY at 1:500 dilution.…”

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confidence: 99%

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Loss of a GPI-Anchored Membrane Protein Aah3p Causes a Defect in Vacuolar Protein Sorting inSchizosaccharomyces pombe

Iwaki

Morita

Tanaka

et al. 2007

Bioscience, Biotechnology, and Biochemistry

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Vacuolar protein sorting receptor in Schizosaccharomyces pombe

Cited by 44 publications

References 62 publications

Vsl1p cooperates with Fsv1p for vacuolar protein transport and homotypic fusion in Schizosaccharomyces pombe

Vsl1p cooperates with Fsv1p for vacuolar protein transport and homotypic fusion in Schizosaccharomyces pombe

Regulation of Ras Localization and Cell Transformation by Evolutionarily Conserved Palmitoyltransferases

Loss of a GPI-Anchored Membrane Protein Aah3p Causes a Defect in Vacuolar Protein Sorting inSchizosaccharomyces pombe

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