González-Párraga Pilar, Hernández José A, Argüelles Juan Carlos
Area de Microbiología, Facultad de Biología, Universidad de Murcia, E-30071 Murcia, Spain.
Yeast. 2003 Oct 30;20(14):1161-9. doi: 10.1002/yea.1029.
In Candida albicans, trehalose plays an essential role as a protector of cell integrity against oxidative challenge. A double homozygous mutant, tps1/tps1, deficient in trehalose synthesis, displayed severe cell mortality when exposed to high H(2)O(2) concentrations, compared with its congenic parental (CAI-4) strain (Alvarez-Peral et al., 2002). We have examined the putative role of a set of well-known antioxidant enzymes as components of the defence mechanism against oxidative challenges. When exposed to mild non-lethal oxidative treatment (0.5 mM H(2)O(2)), a significant induction of catalase, glutathione reductase (GR), and Cu,Zn-superoxide dismutase (SOD) was recorded in tps1/tps1 exponential cultures. However, in CAI-4 cells, subjected to the same conditions, there was only a clear activation of catalase, Mn-SOD and Cu,Zn-SOD activities. The degree of activation was always much more pronounced in the trehalose-deficient mutant than in its wild-type counterpart, except for Mn-SOD activity. After exposure to severe oxidative stress (50 mM H(2)O(2)) only GR and catalase activities increased in tps1/tps1 cultures, whereas in CAI-4 cells GR but not catalase was induced. In both cell strains, 50 mM H(2)O(2) caused inhibition of the Mn- and Cu,Zn-SOD isozymes, this inhibition being more pronounced in tps1/tps1 cells. C. albicans is able to acquire adaptive oxidative tolerance by pretreatment with a low non-stressing concentration of H(2)O(2) before exposure to a drastic oxidative challenge. When these antioxidant activities were measured during the adaptive response, a greater degree of enzymatic antioxidant induction was consistently observed in the tps1/tps1 mutant with respect to the CAI-4 strain. Together with a higher intrinsic sensitivity of tps1/tps1 cells, we suggest that this unexpected increase might be explained in terms of a compensatory mechanism to overcome the lack of endogenous trehalose upon drastic oxidative exposure, although this induction was not sufficient to improve the percentage of cell viability.
在白色念珠菌中,海藻糖作为细胞完整性的保护剂,在抵抗氧化应激方面发挥着重要作用。与同基因亲本(CAI-4)菌株相比,海藻糖合成缺陷的双纯合突变体tps1/tps1在暴露于高浓度H₂O₂时表现出严重的细胞死亡(Alvarez-Peral等人,2002年)。我们研究了一组著名的抗氧化酶作为抗氧化应激防御机制组成部分的假定作用。当暴露于轻度非致死性氧化处理(0.5 mM H₂O₂)时,在tps1/tps1指数生长期培养物中,过氧化氢酶、谷胱甘肽还原酶(GR)和铜锌超氧化物歧化酶(SOD)有显著诱导。然而,在相同条件下的CAI-4细胞中,只有过氧化氢酶、锰超氧化物歧化酶和铜锌超氧化物歧化酶活性明显激活。除了锰超氧化物歧化酶活性外,海藻糖缺陷突变体中的激活程度总是比其野生型对应物更明显。暴露于严重氧化应激(50 mM H₂O₂)后,tps1/tps1培养物中只有GR和过氧化氢酶活性增加,而在CAI-4细胞中,GR被诱导,但过氧化氢酶未被诱导。在两种细胞株中,50 mM H₂O₂均导致锰和铜锌超氧化物歧化酶同工酶的抑制,这种抑制在tps1/tps1细胞中更明显。白色念珠菌能够通过在暴露于剧烈氧化应激之前用低浓度无应激的H₂O₂进行预处理来获得适应性氧化耐受性。当在适应性反应过程中测量这些抗氧化活性时,相对于CAI-4菌株,在tps1/tps1突变体中始终观察到更高程度的酶促抗氧化诱导。结合tps1/tps1细胞更高的内在敏感性,我们认为这种意外增加可能是由于一种补偿机制来克服剧烈氧化暴露时内源性海藻糖的缺乏,尽管这种诱导不足以提高细胞活力百分比。