Poleg Y, Aramayo R, Kang S, Hall J G, Metzenberg R L
Department of Biomolecular Chemistry, University of Wisconsin, Madison 53706, USA.
Mol Gen Genet. 1996 Oct 28;252(6):709-16. doi: 10.1007/BF02173977.
In response to phosphorus limitation, the fungus Neurospora crassa synthesizes a number of enzymes that function to bring more phosphate into the cell. The NUC-2 protein appears to sense the availability of phosphate and transmits the signal downstream to the regulatory pathway. The nuc-2+ gene has been cloned by its ability to restore growth of a nuc-2 mutant under restrictive conditions of high pH and low phosphate concentration. We mapped the cloned gene to the right arm of linkage group II, consistent with the chromosomal position of the nuc-2 mutation as determined by classical genetic mapping. The nuc-2' open reading frame is interrupted by five introns and codes for a protein of 1066 amino acid residues. Its predicted amino acid sequence has high similarity to that of its homolog in Saccharomyces cerevisiae, PHO81. Both proteins contain six ankyrin repeats, which have been implicated in the cyclin-dependent kinase inhibitory activity of PHO81. The phenotypes of a nuc-2 mutant generated by repeat-induced point mutation and of a strain harboring a UV-induced nuc-2 allele are indistinguishable. Both are unable to grow under the restrictive conditions, a phenotype which is to some degree temperature dependent. The nuc-2+ gene is transcriptionally regulated. A 15-fold increase in the level of the nuc-2+ transcript occurs in response to a decrease in exogenous phosphate concentration.
为响应磷限制,真菌粗糙脉孢菌会合成多种酶,这些酶的作用是将更多磷酸盐带入细胞。NUC - 2蛋白似乎能感知磷酸盐的可用性,并将信号传递到下游的调节途径。nuc - 2⁺基因已通过其在高pH和低磷酸盐浓度的限制条件下恢复nuc - 2突变体生长的能力而被克隆。我们将克隆的基因定位到连锁群II的右臂,这与通过经典遗传图谱确定的nuc - 2突变的染色体位置一致。nuc - 2'开放阅读框被五个内含子打断,编码一个由1066个氨基酸残基组成的蛋白质。其预测的氨基酸序列与其在酿酒酵母中的同源物PHO81具有高度相似性。这两种蛋白质都含有六个锚蛋白重复序列,这些重复序列与PHO81的细胞周期蛋白依赖性激酶抑制活性有关。由重复诱导点突变产生的nuc - 2突变体和携带紫外线诱导的nuc - 2等位基因的菌株的表型无法区分。两者在限制条件下都无法生长,这种表型在一定程度上依赖于温度。nuc - 2⁺基因受到转录调控。响应外源磷酸盐浓度的降低,nuc - 2⁺转录本水平增加了15倍。