Blacketer M J, Madaule P, Myers A M
Department of Biochemistry and Biophysics, Iowa State University, Ames 50011.
Mol Cell Biol. 1994 Jul;14(7):4671-81. doi: 10.1128/mcb.14.7.4671-4681.1994.
Saccharomyces cerevisiae mutant E124 was selected in a visual screen based on elongated cell shape. Genetic analysis showed that E124 contains two separate mutations, pps1-1 and elm4-1, each causing a distinct phenotype inherited as a single-gene trait. In rich medium, pps1-1 by itself causes increased doubling time but does not affect cell shape, whereas elm4-1 results in a moderate cell elongation phenotype but does not affect growth rate. Reconstructed elm4-1 pps1-1 double mutants display a synthetic phenotype in rich medium including extreme cell elongation and delayed cell separation, both characteristics of pseudohyphal differentiation. The elm4-1 mutation was shown to act as a dominant factor that potentiates pseudohyphal differentiation in response to general nitrogen starvation in a genetic background in which pseudohyphal growth normally does not occur. Thus, elm4-1 allows recognition of, or response to, a pseudohyphal differentiation signal that results from nitrogen limitation. PPS1 was isolated and shown to be a previously undescribed gene coding for a protein similar in amino acid sequence to phosphoribosylpyrophosphate synthase, a rate-limiting enzyme in the biosynthesis of nucleotides, histidine, and tryptophan. Thus, the pps1-1 mutation may generate a nitrogen limitation signal, which when coupled with elm4-1 results in pseudohyphal growth even in rich medium.
酿酒酵母突变体E124是基于细胞形状拉长在视觉筛选中被挑选出来的。遗传分析表明,E124包含两个独立的突变,pps1-1和elm4-1,每个突变都导致一种作为单基因性状遗传的独特表型。在丰富培养基中,单独的pps1-1会导致倍增时间增加,但不影响细胞形状,而elm4-1会导致中等程度的细胞伸长表型,但不影响生长速率。重建的elm4-1 pps1-1双突变体在丰富培养基中表现出一种合成表型,包括极端的细胞伸长和延迟的细胞分离,这两个都是假菌丝分化的特征。elm4-1突变被证明是一个显性因子,在通常不会发生假菌丝生长的遗传背景下,它能增强对一般氮饥饿的假菌丝分化反应。因此,elm4-1能够识别或响应由氮限制产生的假菌丝分化信号。PPS1被分离出来,结果表明它是一个以前未被描述的基因,编码一种氨基酸序列与磷酸核糖焦磷酸合成酶相似的蛋白质,磷酸核糖焦磷酸合成酶是核苷酸、组氨酸和色氨酸生物合成中的一种限速酶。因此,pps1-1突变可能产生一种氮限制信号,当与elm4-1结合时,即使在丰富培养基中也会导致假菌丝生长。