Cabib E, Sburlati A, Bowers B, Silverman S J
Laboratory of Biochemistry and Metabolism, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892.
J Cell Biol. 1989 May;108(5):1665-72. doi: 10.1083/jcb.108.5.1665.
Previously, we showed that chitin synthase 2 (Chs2) is required for septum formation in Saccharomyces cerevisiae, whereas chitin synthase 1 (Chs1) does not appear to be an essential enzyme. However, in strains carrying a disrupted CHS1 gene, frequent lysis of buds is observed. Lysis occurs after nuclear separation and appears to result from damage to the cell wall, as indicated by osmotic stabilization and by a approximately 50-nm orifice at the center of the birth scar. Lysis occurs at a low pH and is prevented by buffering the medium above pH 5. A likely candidate for the lytic system is a previously described chitinase that is probably involved in cell separation. The chitinase has a very acidic pH optimum and a location in the periplasmic space that exposes it to external pH. Accordingly, allosamidin, a specific chitinase inhibitor, substantially reduced the number of lysed cells. Because the presence of Chs1 in the cell abolishes lysis, it is concluded that damage to the cell wall is caused by excessive chitinase activity at acidic pH, which can normally be repaired through chitin synthesis by Chs1. The latter emerges as an auxiliary or emergency enzyme. Other experiments suggest that both Chs1 and Chs2 collaborate in the repair synthesis of chitin, whereas Chs1 cannot substitute for Chs2 in septum formation.
先前,我们发现几丁质合酶2(Chs2)是酿酒酵母中隔膜形成所必需的,而几丁质合酶1(Chs1)似乎不是一种必需酶。然而,在携带CHS1基因缺失的菌株中,经常观察到芽的裂解。裂解发生在核分离之后,似乎是由细胞壁损伤导致的,渗透稳定作用以及芽痕中心大约50纳米的小孔表明了这一点。裂解在低pH值下发生,通过将培养基缓冲至pH值高于5可防止裂解。裂解系统的一个可能候选者是先前描述的一种几丁质酶,它可能参与细胞分离。该几丁质酶具有非常酸性的最适pH值,并且位于周质空间中,使其暴露于外部pH值。因此,特异性几丁质酶抑制剂阿洛沙米定显著减少了裂解细胞的数量。由于细胞中Chs1的存在消除了裂解现象,因此可以得出结论,细胞壁损伤是由酸性pH值下过量的几丁质酶活性引起的,而这种损伤通常可以通过Chs1的几丁质合成来修复。Chs1作为一种辅助或应急酶出现。其他实验表明,Chs1和Chs2在几丁质的修复合成中协同作用,而Chs1在隔膜形成中不能替代Chs2。