The cell wall of Saccharomyces cerevisiae is an elastic structure that provides osmotic and physical protection and determines the shape of the cell. The inner layer of the wall is largely responsible for the mechanical strength of the wall and also provides the attachment sites for the proteins that form the outer layer of the wall. Here we find among others the sexual agglutinins and the flocculins. The outer protein layer also limits the permeability of the cell wall, thus shielding the plasma membrane from attack by foreign enzymes and membrane-perturbing compounds. The main features of the molecular organization of the yeast cell wall are now known. Importantly, the molecular composition and organization of the cell wall may vary considerably. For example, the incorporation of many cell wall proteins is temporally and spatially controlled and depends strongly on environmental conditions. Similarly, the formation of specific cell wall protein–polysaccharide complexes is strongly affected by external conditions. This points to a tight regulation of cell wall construction. Indeed, all five mitogen-activated protein kinase pathways in bakers’ yeast affect the cell wall, and additional cell wall-related signaling routes have been identified. Finally, some potential targets for new antifungal compounds related to cell wall construction are discussed.

Dynamics of cell wall structure in Saccharomyces cerevisiae.

In this review, we discuss new insights in cell wall architecture and cell wall construction in the ascomycetous yeast Saccharomyces cerevisiae. Transcriptional profiling studies combined with biochemical work have provided ample evidence that the cell wall is a highly adaptable organelle. In particular, the protein population that is anchored to the stress-bearing polysaccharides of the cell wall, and forms the interface with the outside world, is highly diverse. This diversity is believed to play an important role in adaptation of the cell to environmental conditions, in growth mode and in survival. Cell wall construction is tightly controlled and strictly coordinated with progression of the cell cycle. This is reflected in the usage of specific cell wall proteins during consecutive phases of the cell cycle and in the recent discovery of a cell wall integrity checkpoint. When the cell is challenged with stress conditions that affect the cell wall, a specific transcriptional response is observed that includes the general stress response, the cell wall integrity pathway and the calcineurin pathway. This salvage mechanism includes increased expression of putative cell wall assemblases and some potential cross-linking cell wall proteins, and crucial changes in cell wall architecture. We discuss some more enzymes involved in cell wall construction and also potential inhibitors of these enzymes. 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.

Cell wall construction in Saccharomyces cerevisiae.

The chemical composition of many fungal cell walls is known, but we have not understood the interactions of the various macromolecules nor the assembly processes. The biochemistry and molecular genetics of biosynthesis have been comprehensively reviewed ([9][1], [42][2]), but recent results obtained

Cell Wall Architecture in Yeast: New Structure and New Challenges

https://pure.uva.nl/ws/files/4058964/2082_21713y.pdf

Architecture of the yeast cell wall. Beta(1-->6)-glucan interconnects mannoprotein, beta(1-->)3-glucan, and chitin.

Chitin is the (1→4)-β-linked homopolymer of N-acetyl-D-glucosamine (Fig. 1). The individual polymer chains can be thought of as helices, as each sugar unit is inverted with respect to its neighbors. This leads to their stabilization as rigid ribbons by 03—H … 05 and 06—H … 07 hydrogen bonds. The commonest form of chitin is α-chitin. Its unit cell is of two N,N‵-diacetylchitobiose units of two chains in an antiparallel arrangement. Thus, adjacent polymer chains run in opposite directions, held together by 06—H … 06 hydrogen bonds, and the chains are held in sheets by 07 … H—N hydrogen bonds (Minke and Blackwell, 1978). This gives a statistical mixture of CH2OH orientations, equivalent to half oxygens on each residue, each forming inter- and intramolecular hydrogen bonds. This results in two types of amide groups; all are involved in the interchain C=O … H—N bonds, while half of the groups also serve as acceptors for 06—H … O=C intramolecular bonds. This extensive intermolecular hydrogen bonding leads to a very stable structure, the individual polymer chains eventually giving rise to microfibrils if allowed to crystallize (Gooday, 1983). A less common form of chitin is β-chitin, in which the unit cell is of one N,N‵diacetylchitobiose unit, giving a polymer stabilized as a rigid ribbon, as for α-chitin, by 03—H … 05 intramolecular bonds (Gardner and Blackwell, 1975). Chains are then held together in sheets by C=O … H—N hydrogen bonding of the amide groups and by the CH2OH side chains, forming intersheet hydrogen bonds to the carbonyl oxygens on the next chains (06—H … 07). This gives a structure of parallel poly-N-acetylglucosamine chains with no intersheet hydrogen bonds.

The Ecology of Chitin Degradation

Pulse-chase experiments with [14C]glucose demonstrated that in the cell wall of wild-type Saccharomyces cerevisiae alkali-soluble (1-3)-beta-glucan serves as a precursor for alkali-insoluble (1-3)-beta-glucan. The following observations support the notion that the insolubilization of the glucan is caused by linkage to chitin: (i) degradation of chitin by chitinase completely dissolved the glucan, and (ii) disruption of the gene for chitin synthase 3 prevented the formation of alkali-insoluble glucan. These cells, unable to form a glucan-chitin complex, were highly vulnerable to hypo-osmotic shock indicating that the linkage of the two polymers significantly contributes to the mechanical strength of the cell wall. Conversion of alkali-soluble glucan into alkali-insoluble glucan occurred both early and late during budding and also in the ts-mutant cdc24-1 in the absence of bud formation.

The linkage of (1-3)-beta-glucan to chitin during cell-wall assembly in saccharomyces-cerevisiae

Chitin is a structural component present in the wall of all fungi except the Oomycetes where cellulose plays a similar role. It is usually considered to be present as microfibrils, consisting of crystalline chitin embedded in an amorphous matrix (1), because most electronmicroscopical and X-ray diffraction studies show microfibrillar and crystalline chitin. However these results were usually obtained after drastic chemical treat¬ments of the wall and induction of crystallisation and microfibril formation by these treatments remains a possibility (2).

C. A. Vermeulen

Papers

The linkage of (1-3)-beta-glucan to chitin during cell-wall assembly in saccharomyces-cerevisiae

The Role of Chitin in Hyphal Morphogenesis