Gillmor C Stewart, Poindexter Patricia, Lorieau Justin, Palcic Monica M, Somerville Chris
Carnegie Institution, Department of Plant Biology, Stanford, CA 94305, USA.
J Cell Biol. 2002 Mar 18;156(6):1003-13. doi: 10.1083/jcb.200111093.
Novel mutations in the RSW1 and KNOPF genes were identified in a large-scale screen for mutations that affect cell expansion in early Arabidopsis embryos. Embryos from both types of mutants were radially swollen with greatly reduced levels of crystalline cellulose, the principal structural component of the cell wall. Because RSW1 was previously shown to encode a catalytic subunit of cellulose synthase, the similar morphology of knf and rsw1-2 embryos suggests that the radially swollen phenotype of knf mutants is largely due to their cellulose deficiency. Map-based cloning of the KNF gene and enzyme assays of knf embryos demonstrated that KNF encodes alpha-glucosidase I, the enzyme that catalyzes the first step in N-linked glycan processing. The strongly reduced cellulose content of knf mutants indicates that N-linked glycans are required for cellulose biosynthesis. Because cellulose synthase catalytic subunits do not appear to be N glycosylated, the N-glycan requirement apparently resides in other component(s) of the cellulose synthase machinery. Remarkably, cellular processes other than extracellular matrix biosynthesis and the formation of protein storage vacuoles appear unaffected in knf embryos. Thus in Arabidopsis cells, like yeast, N-glycan trimming is apparently required for the function of only a small subset of N-glycoproteins.
在一项大规模筛选中,研究人员在拟南芥早期胚胎中发现了影响细胞扩张的突变,其中鉴定出了RSW1和KNOPF基因的新突变。这两种突变体的胚胎均呈径向肿胀,细胞壁的主要结构成分结晶纤维素水平大幅降低。由于之前研究表明RSW1编码纤维素合酶的一个催化亚基,knf和rsw1-2胚胎相似的形态表明,knf突变体的径向肿胀表型很大程度上是由于其纤维素缺乏所致。对KNF基因进行图位克隆以及对knf胚胎进行酶活性分析表明,KNF编码α-葡糖苷酶I,该酶催化N-连接聚糖加工的第一步。knf突变体中纤维素含量大幅降低,这表明N-连接聚糖是纤维素生物合成所必需的。由于纤维素合酶催化亚基似乎不进行N-糖基化,N-聚糖的需求显然存在于纤维素合酶机制的其他成分中。值得注意的是,除了细胞外基质生物合成和蛋白质储存液泡形成之外的细胞过程在knf胚胎中似乎未受影响。因此,在拟南芥细胞中,与酵母一样,N-聚糖修剪显然仅对一小部分N-糖蛋白的功能是必需的。