Frans M Klis

..Finally, we use both biochemical and genomic data to infer that the architectural principles used by S. cerevisiae to build its cell wall are also used by many other ascomycetous yeasts and also by some mycelial ascomycetous fungi...

..The wall proteome of the opportunistic human pathogen Candida albicans amply illustrates these properties. Finally, we discuss the relevance of our insights for the identification of new vaccine candidates...

..We conclude that covalently linked CWPs of C. albicans play a crucial role in fitness and virulence and that their expression is tightly controlled...

..We also discuss the hybrid nature of many wall proteins and the frequent occurrence of families of wall proteins with a common multi-domain structure. Finally, some quantitative data regarding wall proteins are presented...

..Similarly, biotinylation of cell surface proteins should be carried out at low temperatures in the absence of dimethylsulphoxide...

..Addition of the iron chelator ferrozine at saturating levels of O(2) resulted in higher cell wall levels of Hwp1 and Rbt5, suggesting that the responses to hypoxic conditions and iron restriction are related...

..We propose that the large repertoire of adhesin(-like) genes of C. glabrata contributes to its adaptability and virulence...

..Taken together, our data indicate that cell wall stress results in activation of various signalling pathways including the cell wall integrity pathway...

..cerevisiae and Candida albicans, the amount of carbohydrate added to the proteins was found to be two- to three-fold decreased, consistent with earlier reported differences in outer chain N-glycosylation...

..This is, to our knowledge, the first systematic, quantitative analysis of the changes in the wall proteome of C. albicans upon hyphal induction. Finally, we propose new wall-protein-derived candidates for vaccine development...

..Therefore, it will probably be equally effective in many other fungi...

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..These results demonstrate that ambient pH strongly affects the wall proteome of C. albicans, show that our quantitative approach can give detailed insights into the dynamics of the wall proteome, and point to potential vaccine targets...

..The levels of Crh1p, Crh2p, Ecm33p, Gas5p, Pst1p and Pir3p increased about three- to fivefold, whereas the level of Scw4p was significantly decreased. We propose that our methods are widely applicable to other fungi...

..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...

..3p, Pir1p, Pga29p, Rbt5p, and Ssr1p) are unknown. These results indicate that a substantial number of the covalently linked CWPs of C. albicans are actively involved in cell wall remodeling and expansion and in host-pathogen interactions...

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..In summary, the number of identified secretory proteins in the growth medium has been substantially extended, and growth conditions strongly affect the composition of the secretome...

..Finally, we discuss the implications of our results for combinatorial drug design...

..nidulans, including ten predicted GPI proteins, most of which belong to carbohydrate-active enzyme families that are also found in the walls of ascomycetous yeasts...

..pombe and N. crassa are homologous to known or putative GPI proteins from other fungi. We conclude that our algorithm is selective and can also be used for GPI protein identification in other fungi...

..We propose that surface stresses lead to a conserved cell wall remodeling response that is mainly governed by Mkc1 and is characterized by chitin reinforcement of the wall and the expression of remedial wall remodeling enzymes...

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..We propose that global fitness analysis of yeast in combination with T-profiler is a powerful tool to identify specific cellular processes and pathways that are required for survival under stress conditions...

..Extracts from rho2, rho3, rho4 and rho5 null mutants had wild-type in vitro activity. In contrast, Rho1p overproduction led to increased in vitro synthesis, implicating Rho1p in the regulation of (1 --> 6)-beta-D-glucan synthesis...

..We present evidence that the cell wall of mycelial species of the Ascomycotina is similarly organized and contains glycoproteins with comparable functions...

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..We also present candidate proteins from the secretome for clinical marker development...

..Overall, we discuss the importance of the secretome for virulence and suggest potential targets for better and faster diagnostic methods...

..It is proposed that Kic1 affects cell wall construction in multiple ways and in particular in regulating 1,6-beta-glucan levels in the wall...

..We propose that the transcriptional response to chitosan may be representative of other plasma membrane-perturbing compounds...

..Finally, some potential targets for new antifungal compounds related to cell wall construction are discussed...

..Our results indicate that galactofuranose formation is important for fungal cell wall biosynthesis and represents an attractive target for the development of antifungals...

..A CwpA-specific antiserum was raised and in combination with fractionation experiments, we show that this protein was abundantly present as an HF-extractable protein in the cell wall of A. niger...

..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...

..niger, but was also observed in Penicillium chrysogenum. We propose that this response to cell wall stress commonly occurs in filamentous fungi...

..In conclusion, this work establishes the role of UTR2, CRH11, and CRH12 in cell wall organization and integrity...

..We propose that such a cell wall salvage mechanism is wide spread in filamentous fungi...

..niger. Noteworthy is the large number of major facilitator superfamily transporters and fungal zinc binuclear cluster transcription factors, and the presence of putative gene clusters for fumonisin and ochratoxin A synthesis...

..The protocol can be completed in 4-6 h (excluding the incubation time required for fungal growth)...

..These observations suggest that cell wall stress in fungi may generally lead to activation of the chitin biosynthetic pathway...

..This mature form of cell wall alpha-glucan is essential for fission-yeast morphogenesis...

..These aspects of fungal adhesion exemplify the impressive phenotypic plasticity of yeasts, allowing them to adapt quickly to stressful environments and exploit new opportunities...

..We propose that the cell wall proteins thus identified are important for pathogenicity...