Pga1 Is an Essential Component of Glycosylphosphatidylinositol-Mannosyltransferase II of<i>Saccharomyces cerevisiae</i>

Sato, Keisuke; Noda, Yoichi; Yoda, Koji

doi:10.1091/mbc.e07-03-0258

Cited by 24 publications

(24 citation statements)

References 46 publications

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“…Anti-HA (12CA5, Roche Diagnostics) monoclonal antibody was purchased. Rabbit polyclonal antibody to Gas1 was described previously (15). For Western blotting, anti-Kre6 rabbit antiserum was used at a dilution of 1/500, and the other antisera were used at a dilution of 1/1000.…”

Section: Methodsmentioning

confidence: 99%

Kre6 Protein Essential for Yeast Cell Wall β-1,6-Glucan Synthesis Accumulates at Sites of Polarized Growth

Kurita

Noda

Такаги

et al. 2011

Journal of Biological Chemistry

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Saccharomyces cerevisiae Kre6 is a type II membrane protein with amino acid sequence homology with glycoside hydrolase and is essential for ␤-1,6-glucan synthesis as revealed by the mutant phenotype, but its biochemical function is still unknown. The localization of Kre6, determined by epitope tagging, is a matter of debate. We raised anti-Kre6 rabbit antiserum and examined the localization of Kre6 and its tagged protein by immunofluorescence microscopy, subcellular fractionation in sucrose density gradients, and immunoelectron microscopy. Integration of the results indicates that the majority of Kre6 is in the endoplasmic reticulum; however, a small but significant portion is also present in the secretory vesiclelike compartments and plasma membrane. Kre6 in the latter compartments is observed as strong signals that accumulate at the sites of polarized growth by immunofluorescence. The truncated Kre6 without the N-terminal 230-amino acid cytoplasmic region did not show this polarized accumulation and had a severe defect in ␤-1,6-glucan synthesis. This is the first evidence of a ␤-1,6-glucan-related protein showing the polarized membrane localization that correlates with its biological function.

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Section: Methodsmentioning

confidence: 99%

Kre6 Protein Essential for Yeast Cell Wall β-1,6-Glucan Synthesis Accumulates at Sites of Polarized Growth

Kurita

Noda

Такаги

et al. 2011

Journal of Biological Chemistry

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“…Addition of a1,6-linked Man-2 requires catalytic Gpi18 Kang et al 2005) and Pga1 (Sato et al 2007), which form a complex (Sato et al 2007). Gpi18-deficient cells accumulate both a Man 1 -GPI with Etn-P esterified to its Man and an unmodified Man 1 -GPI, suggesting that GPI-Man-T-II can use either as acceptor Scarcelli et al 2012).…”

Section: Gpi Anchoringmentioning

confidence: 99%

Architecture and Biosynthesis of the Saccharomyces cerevisiae Cell Wall

2012

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The wall gives a Saccharomyces cerevisiae cell its osmotic integrity; defines cell shape during budding growth, mating, sporulation, and pseudohypha formation; and presents adhesive glycoproteins to other yeast cells. The wall consists of b1,3-and b1,6-glucans, a small amount of chitin, and many different proteins that may bear N-and O-linked glycans and a glycolipid anchor. These components become cross-linked in various ways to form higher-order complexes. Wall composition and degree of cross-linking vary during growth and development and change in response to cell wall stress. This article reviews wall biogenesis in vegetative cells, covering the structure of wall components and how they are cross-linked; the biosynthesis of N-and O-linked glycans, glycosylphosphatidylinositol membrane anchors, b1,3-and b1,6-linked glucans, and chitin; the reactions that cross-link wall components; and the possible functions of enzymatic and nonenzymatic cell wall proteins. TABLE OF CONTENTS Abstract 775Introduction 777

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“…The localization of Aur1-3HA and Kre2-3HA was observed by the indirect immunofluorescence as described previously (Sato et al, 2007) except that anti-HA antibody and Alexa 568-conjugated goat antibody to mouse immunoglobulin G were used as the primary antibody and the secondary antibody, respectively.…”

Section: Indirect Immunofluorescencementioning

confidence: 99%

“…We have tried to assign functions to such essential proteins that are integrated into membranes by our strategy of using temperature-sensitive mutants. To date, we have reported the characterization of two previously uncharacterized endoplasmic reticulum (ER) proteins Pga1 and Keg1 (Nakamata et al, 2007;Sato et al, 2007). YDR367w is one of such uncharacterized essential gene, encoding a 221-amino acid polypeptide with four predicted transmembrane domains.…”

Section: Introductionmentioning

confidence: 99%

Kei1: A Novel Subunit of Inositolphosphorylceramide Synthase, Essential for Its Enzyme Activity and Golgi Localization

Sato

Noda

Yoda

2009

MBoC

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Fungal sphingolipids have inositol-phosphate head groups, which are essential for the viability of cells. These head groups are added by inositol phosphorylceramide (IPC) synthase, and AUR1 has been thought to encode this enzyme. Here, we show that an essential protein encoded by KEI1 is a novel subunit of IPC synthase of Saccharomyces cerevisiae. We find that Kei1 is localized in the medial-Golgi and that Kei1 is cleaved by Kex2, a late Golgi processing endopeptidase; therefore, it recycles between the medial-and late Golgi compartments. The growth defect of kei1-1, a temperaturesensitive mutant, is effectively suppressed by the overexpression of AUR1, and Aur1 and Kei1 proteins form a complex in vivo. The kei1-1 mutant is hypersensitive to aureobasidin A, a specific inhibitor of IPC synthesis, and the IPC synthase activity in the mutant membranes is thermolabile. A part of Aur1 is missorted to the vacuole in kei1-1 cells. We show that the amino acid substitution in kei1-1 causes release of Kei1 during immunoprecipitation of Aur1 and that Aur1 without Kei1 has hardly detectable IPC synthase activity. From these results, we conclude that Kei1 is essential for both the activity and the Golgi localization of IPC synthase.

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Pga1 Is an Essential Component of Glycosylphosphatidylinositol-Mannosyltransferase II ofSaccharomyces cerevisiae

Cited by 24 publications

References 46 publications

Kre6 Protein Essential for Yeast Cell Wall β-1,6-Glucan Synthesis Accumulates at Sites of Polarized Growth

Kre6 Protein Essential for Yeast Cell Wall β-1,6-Glucan Synthesis Accumulates at Sites of Polarized Growth

Architecture and Biosynthesis of the Saccharomyces cerevisiae Cell Wall

Kei1: A Novel Subunit of Inositolphosphorylceramide Synthase, Essential for Its Enzyme Activity and Golgi Localization

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