Liu Xiaoguang, Hu Guowu, Panepinto John, Williamson Peter R
Section of Infectious Diseases, Department of Medicine, University of Illinois at Chicago, College of Medicine, Chicago, IL, USA.
Mol Microbiol. 2006 Sep;61(5):1132-46. doi: 10.1111/j.1365-2958.2006.05299.x.
Previous studies have demonstrated an important role for the vacuole in the virulence of the fungus Cryptococcus and studies in yeast have implicated the vacuolar protein Vps41 in copper loading of proteins such as iron transporters. However, our studies found that a cryptococcal vps41Delta strain displayed wild-type growth on media containing iron and copper chelators and normal activity of the copper-containing virulence factor laccase as well as almost normal growth at 37 degrees C and wild-type production of the virulence factor capsule. Despite these attributes, the vps41Delta mutant strain showed a dramatic attenuation of virulence in mice and co-incubation of mutant cells with the macrophage cell line, J774.16, resulted in a dramatic loss in viability of the vps41Delta mutant strain at 10 h compared with wild-type and complemented strains. Closer examination revealed that the vps41Delta mutant displayed a dramatic loss in viability after nutrient starvation which was traced to a failure to undergo G2 arrest, but there was no defect in the formation of autophagic or proteolytic vesicles. Our results indicate that VPS41 plays a key role in regulating starvation response in this pathogenic organism and that defects in cell cycle arrest are associated with attenuated pathogenic fitness in mammalian hosts.
先前的研究已经证明液泡在新型隐球菌的毒力中发挥重要作用,并且在酵母中的研究表明液泡蛋白Vps41参与了诸如铁转运蛋白等蛋白质的铜装载过程。然而,我们的研究发现,新型隐球菌vps41Δ菌株在含有铁和铜螯合剂的培养基上呈现野生型生长,含铜毒力因子漆酶具有正常活性,在37℃时生长几乎正常,毒力因子荚膜的产生也为野生型。尽管具有这些特性,但vps41Δ突变菌株在小鼠中的毒力显著减弱,并且与巨噬细胞系J774.16共同孵育时,与野生型和互补菌株相比,vps41Δ突变菌株在10小时时活力急剧丧失。进一步检查发现,vps41Δ突变体在营养饥饿后活力急剧丧失,这可追溯到无法进入G2期停滞,但自噬或蛋白水解囊泡的形成没有缺陷。我们的结果表明,VPS41在调节这种致病生物的饥饿反应中起关键作用,并且细胞周期停滞缺陷与在哺乳动物宿主中的致病适应性减弱有关。