Mitsuzawa Hiroshi
Department of Applied Biological Sciences, Nihon University College of Bioresource Sciences, Fujisawa, Kanagawa 252-8510, Japan.
Genes Cells. 2006 Oct;11(10):1183-95. doi: 10.1111/j.1365-2443.2006.01014.x.
Ammonium is an important source of nitrogen for many microorganisms, including yeast, and its availability also has substantial effects on the nitrogen metabolism and development of yeast cells. Three ammonium transporter genes of the fission yeast Schizosaccharomyces pombe, named amt1, amt2, and amt3, were identified on the basis of amino acid sequence similarity to members of the ammonium transporter/methylammonium permease (Amt/Mep) family. A series of strains were constructed that carry all combinations of amt deletion (amt delta) mutations, and tested for growth on low ammonium and resistance to the toxic ammonium analog methylammonium. The amt1 delta and amt2 delta single mutants had different growth defects, and the amt1 delta amt2 delta double mutant displayed a much more severe growth defect on < or = 5 mM ammonium. All single mutants exhibited methylammonium resistance but to different extents: amt2 delta was the most resistant and amt3 delta was the least. These results suggest that the amt genes encode functional transporters with distinct uptake properties. In response to ammonium limitation, the wild-type strain isogenic to the amt delta mutants underwent filamentous growth underneath the surface of solid medium. No such filamentous invasive growth, however, was observed for the amt1 delta mutant, indicating that Amt1 transporter is required for ammonium limitation-induced filamentous invasive growth.
铵是包括酵母在内的许多微生物的重要氮源,其可利用性对酵母细胞的氮代谢和发育也有重大影响。基于与铵转运蛋白/甲基铵通透酶(Amt/Mep)家族成员的氨基酸序列相似性,在裂殖酵母粟酒裂殖酵母中鉴定出三个铵转运蛋白基因,分别命名为amt1、amt2和amt3。构建了一系列携带amt缺失(amtΔ)突变所有组合的菌株,并检测它们在低铵条件下的生长情况以及对有毒铵类似物甲基铵的抗性。amt1Δ和amt2Δ单突变体有不同的生长缺陷,amt1Δamt2Δ双突变体在≤5 mM铵的条件下表现出更严重的生长缺陷。所有单突变体都表现出对甲基铵的抗性,但程度不同:amt2Δ抗性最强,amt3Δ抗性最弱。这些结果表明,amt基因编码具有不同摄取特性的功能性转运蛋白。为响应铵限制,与amtΔ突变体同基因的野生型菌株在固体培养基表面下进行丝状生长。然而,在amt1Δ突变体中未观察到这种丝状侵袭性生长,这表明铵限制诱导的丝状侵袭性生长需要Amt1转运蛋白。