In situ localization of β-glucans in the cell wall of Schizosaccharomyces pombe

被引:111
作者
Humbel, BM
Konomi, M
Takagi, T
Kamasawa, N
Ishijima, SA
Osumi, M
机构
[1] Japan Womens Univ, Fac Sci, Dept Chem & Biol Sci, Bunkyo Ku, Tokyo 1128681, Japan
[2] Japan Womens Univ, Grad Sch Sci, Div Maternal & Biol Sci, Bunkyo Ku, Tokyo 1128681, Japan
关键词
Schizosaccharomyces pombe; cell wall; beta-glucan; cell division; electron microscopy; high-pressure freezing; freeze-substitution; immunogold labelling;
D O I
10.1002/yea.694
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The chemical composition of the cell wall of Sz-. pombe is known as beta -1,3-glucan, beta -1,6-glucan, alpha -1,3-glucan and alpha -galactomannan; however, the three-dimensional interactions of those macromolecules have not yet been clarified. Transmission electron microscopy reveals a three-layered structure: the outer layer is electron-dense, the adjacent layer is less dense, and the third layer bordering the cell membrane is dense. In intact cells of Sc. pombe, the high-resolution scanning electron microscope reveals a surface completely filled with alpha -galactomannan particles. To better understand the organization of the cell wall and to complement our previous studies, we set out to locate the three different types of beta -glucan by immuno-electron microscopy. Our results suggest that the less dense layer of the cell wall contains mainly beta -1,6-branched beta -1,3-glucan. Occasionally a line of gold particles can be seen, labelling fine filaments radiating from the cell membrane to the alpha -galactomannan layer, suggesting that some of the radial filaments contain beta -1,6-branched beta -1,3-glucan. beta -1,6-glucan is preferentially located underneath the alpha -galactomannan layer. Linear beta -1,3-glucan is exclusively located in the primary septum of dividing cells. beta -1,6-glucan only labels the secondary septum and does not co-localize with linear beta -1,3-glucan, while beta -1,6-branched beta -1,3-glucan is present in both septa. Linear beta -1,3-glucan is present from early stages of septum formation and persists until the septum is completely formed; then just before cell division the label disappears. From these results we suggest that linear beta -1,3-glucan is involved in septum formation and perhaps the separation of the two daughter cells. In addition, rye frequently found beta -1,6-glucan label on the Golgi apparatus, on small vesicles and underneath the cell membrane. These results give fresh evidence for the hypothesis that P-1,6-glucan is synthesized in the endoplasmic reticulum-Golgi system and exported to the cell membrane. Copyright (C) 2001 John Wiley & Sons, Ltd.
引用
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页码:433 / 444
页数:12
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