Covalent association of beta-1,3-glucan with beta-1,6-glucosylated mannoproteins in cell walls of Candida albicans

Kapteyn, J.C.; Montijn, R.C.; Dijkgraaf, Gerrit J.P.; Ende, H. van den; Klis, Frans M.

doi:10.1128/jb.177.13.3788-3792.1995

Cited by 103 publications

(99 citation statements)

References 26 publications

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“…on April 27, 2019 by guest http://jb.asm.org/ results provide experimental evidence for the proposal that ␤1,6-glucan plays a functional role in anchoring glucomannoprotein in the cell wall, probably by covalently cross-linking the protein to the extracellular ␤1,3-glucan matrix (22,34). How and where the ␤1,6-glucan side chain is attached onto the protein is not clear, although several possibilities, including that the ␤1,6-glucan is linked to the glycosylphosphatidylinositol anchor (8,21,28,54), have been suggested.…”

Section: Vol 178 1996mentioning

confidence: 81%

CWH41 encodes a novel endoplasmic reticulum membrane N-glycoprotein involved in beta 1,6-glucan assembly

et al. 1996

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CWH41 encodes a novel type II integral membrane N-glycoprotein located in the endoplasmic reticulum. Disruption of the CWH41 gene leads to a K1 killer toxin-resistant phenotype and a 50% reduction in the cell wall ␤1,6-glucan level. CWH41 also displays strong genetic interactions with KRE1 and KRE6, two genes known to be involved in the ␤1,6-glucan biosynthetic pathway. The cwh41⌬ kre6⌬ double mutant is nonviable; and the cwh41⌬ kre1⌬ double mutation results in strong synergistic defects, with a severely slow-growth phenotype, a 75% reduction in ␤1,6-glucan level, and the secretion of a cell wall glucomannoprotein, Cwp1p. These results provide strong genetic evidence indicating that Cwh41p plays a functional role, possibly as a new synthetic component, in the assembly of cell wall ␤1,6-glucan.

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Section: Vol 178 1996mentioning

confidence: 81%

CWH41 encodes a novel endoplasmic reticulum membrane N-glycoprotein involved in beta 1,6-glucan assembly

et al. 1996

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“…Cell walls of C. glabrata were isolated using a method that was previously described for C. albicans (Kapteyn et al, 1995;De Groot et al, 2004). Briefly, yeast cells were grown to exponential phase, washed and homogenized using glass beads in a FastPrep Instrument FP120 (Bio 101).…”

Section: Methodsmentioning

confidence: 99%

Systematic identification in silico of covalently bound cell wall proteins and analysis of protein-polysaccharide linkages of the human pathogen Candida glabrata

Weig

2004

Microbiology

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Candida glabrata is an important cause of systemic candidiasis in humans. This paper reports a systematic analysis of the putative glycosylphosphatidylinositol-modified (GPI) proteins of C. glabrata, a large part of which are covalently bound to the cell wall glucan network and the remainder of which are retained in the plasma membrane, and of cell wall proteins (CWPs) which are covalently bound in a mild-alkali-sensitive manner. In silico genomic analysis revealed 106 putative GPI proteins. Fifty-one of these GPI proteins could be categorized as adhesive proteins, potentially implicated in fungus-host interactions or biofilm formation during the development of fungal infections. Eleven proteins belonged to well-known GPI protein families of glycoside hydrolases, probably involved in cell wall expansion and remodelling during growth. Other identified GPI proteins included phospholipases, aspartic proteases, homologues of ScEcm33p and ScKre1p, and structural CWPs. Interestingly, the GPI algorithm predicted three orthologues of an abundant CWP in S. cerevisiae, Cwp1p, which is absent in Candida albicans. To evaluate the in silico predictions, isolated cell walls were extracted using HF-pyridine, which specifically cleaves phosphodiester bonds, to release GPI-CWPs. Immunological analysis of the extract using one-dimensional SDS-PAGE and anti-ScCwp1p antiserum indicated the presence of a Cwp1p homologue in C. glabrata cell walls. Further analysis by two-dimensional gel electrophoresis and electrospray ionization tandem mass spectrometry (ESI-MS/MS) confirmed the presence of two of the predicted Cwp1p proteins, Cwp1.1p and Cwp1.2p. Crh1p, a putative 1,3-b-glucan remodelling enzyme, was also identified. In silico genomic analysis further revealed five putative Pir proteins (Pir1-5p) and five members of the Bgl2 glycoside hydrolase family 17, belonging to a class of putative CWPs that can be extracted with NaOH. Immunological analysis of mild-alkali-extracted CWPs showed the presence of a ScPir2p homologue. Together, these experimental data and in silico predictions represent the first systematic analysis of the C. glabrata cell wall proteome.

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“…The precipitate was then pelleted at 10,000 ϫ g and washed extensively with 80% (vol/vol) acetone. Meanwhile, the cells were washed and homogenized as described previously (Montijn et al, 1994;Kapteyn et al, 1995), and cell walls were spun down at 3000 ϫ g. The resulting supernatant was precipitated with DOC and TCA as described above ("cytosolic fraction"). Meanwhile, the pellet from the 3000 ϫ g spin was washed extensively with 1 M NaCl and then boiled twice for 10 min in 2% SDS, 100 mM EDTA, 40 mM ␤-mercaptoethanol, and 50 mM Tris-HCl, pH 7.8, to solubilize noncovalently linked cell wall proteins and proteins from the plasma membrane and other membranous compartments .…”

Section: Cell Fractionation Cell Wall Digestion and Western Blottingmentioning

confidence: 99%

Role of Cell Cycle-regulated Expression in the Localized Incorporation of Cell Wall Proteins in Yeast

Smits

Schenkman

Brul

et al. 2006

MBoC

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The yeast cell wall is an essential organelle that protects the cell from mechanical damage and antimicrobial peptides, participates in cell recognition and adhesion, and is important for the generation and maintenance of normal cell shape. We studied the localization of three covalently bound cell wall proteins in Saccharomyces cerevisiae. Tip1p was found only in mother cells, whereas Cwp2p was incorporated in small-to-medium-sized buds. When the promoter regions of TIP1 and CWP2 (responsible for transcription in early G 1 and S/G 2 phases, respectively) were exchanged, the localization patterns of Tip1p and Cwp2p were reversed, indicating that the localization of cell wall proteins can be completely determined by the timing of transcription during the cell cycle. The third protein, Cwp1p, was incorporated into the birth scar, where it remained for several generations. However, we could not detect any role of Cwp1p in strengthening the birth scar wall or any functional interaction with the proteins that mark the birth scar pole as a potential future budding site. Promoter-exchange experiments showed that expression in S/G 2 phase is necessary but not sufficient for the normal localization of Cwp1p. Studies of mutants in which septum formation is perturbed indicate that the normal asymmetric localization of Cwp1p also depends on the normal timing of septum formation, composition of the septum, or both. INTRODUCTIONThe establishment and maintenance of asymmetry are of vital importance for the growth, differentiation, and morphogenesis of cells and organisms. For example, the yeast Saccharomyces cerevisiae grows asymmetrically, producing a bud that becomes the daughter cell (reviewed by Lew and Reed, 1995; Pruyne and Bretscher 2000a,b). In the G 1 phase of the cell cycle, a bud site is selected in a mating type-dependent manner. After the cell cycle-commitment point START, proteins required for bud initiation are recruited to this site, and a bud is formed in a process that depends largely on the delivery of vesicles containing new cell surface material by the polarized actin cytoskeleton. Growth occurs exclusively in the bud, first preferentially at the tip and later isotropically, until the time of cytokinesis. The actin cytoskeleton and cell surface growth are then redirected toward the mother-bud neck, where a chitinous primary septum is formed, followed by the deposition of secondary septa on both sides of the primary septum. Cells then separate by partial digestion of the primary septum, and a new cell cycle is initiated.The cell wall of yeast is essential for growth and is involved in the establishment and maintenance of asymmetry, as illustrated by the loss of both buds and polarized localization of cytoskeletal proteins when the cell wall is removed (Reck-Peterson et al., 1999). The cell wall is comprised of glucans, mannoproteins, and a small amount of chitin (reviewed by Orlean, 1997;Smits et al., 2001;Klis et al., 2006). The mannoproteins can be divided into four classes: 1) noncovalently linked prote...

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Covalent association of beta-1,3-glucan with beta-1,6-glucosylated mannoproteins in cell walls of Candida albicans

Cited by 103 publications

References 26 publications

CWH41 encodes a novel endoplasmic reticulum membrane N-glycoprotein involved in beta 1,6-glucan assembly

CWH41 encodes a novel endoplasmic reticulum membrane N-glycoprotein involved in beta 1,6-glucan assembly

Systematic identification in silico of covalently bound cell wall proteins and analysis of protein-polysaccharide linkages of the human pathogen Candida glabrata

Role of Cell Cycle-regulated Expression in the Localized Incorporation of Cell Wall Proteins in Yeast

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