El-Ganiny Amira M, Sanders David A R, Kaminskyj Susan G W
Department of Biology, University of Saskatchewan, 112 Science Place, Saskatoon, SK, Canada S7N 5E2.
Fungal Genet Biol. 2008 Dec;45(12):1533-42. doi: 10.1016/j.fgb.2008.09.008. Epub 2008 Oct 2.
Growing resistance to current anti-fungal drugs is spurring investigation of new targets, including those in fungal wall metabolism. Galactofuranose (Galf) is found in the cell walls of many fungi including Aspergillus fumigatus, which is currently the most prevalent opportunistic fungal pathogen in developed countries, and A. nidulans, a closely-related, tractable model system. UDP-galactopyranose mutase (UGM) converts UDP-galactopyranose into UDP-Galf prior to incorporation into the fungal wall. We deleted the single-copy UGM sequence (AN3112.4, which we call ugmA) from an A. nidulans nkuADelta strain, creating ugmADelta. Haploid ugmADelta strains were able to complete their asexual life cycle, showing that ugmA is not essential. However, ugmADelta strains had compact colonial growth, which was associated with substantially delayed and abnormal conidiation. Compared to a wildtype morphology strain, ugmADelta strains had aberrant hyphal morphology, producing wide, uneven, highly-branched hyphae, with thick, relatively electron-dense walls as visualized by transmission electron microscopy. These effects were partially remediated by growth on high osmolarity medium, or on medium containing 10 microg/mL Calcofluor, consistent with Galf being important in cell wall structure and/or function.
目前抗真菌药物的耐药性不断增加,这促使人们对新的靶点进行研究,包括那些参与真菌细胞壁代谢的靶点。半乳呋喃糖(Galf)存在于许多真菌的细胞壁中,包括烟曲霉(目前在发达国家中最常见的机会性真菌病原体)和构巢曲霉(一种密切相关的、易于处理的模型系统)。尿苷二磷酸 - 吡喃半乳糖变位酶(UGM)在将其掺入真菌细胞壁之前,将尿苷二磷酸 - 吡喃半乳糖转化为尿苷二磷酸 - Galf。我们从构巢曲霉nkuAΔ菌株中删除了单拷贝的UGM序列(AN3112.4,我们称之为ugmA),从而创建了ugmAΔ。单倍体ugmAΔ菌株能够完成其无性生命周期,这表明ugmA并非必需。然而,ugmAΔ菌株的菌落生长紧凑,这与分生孢子形成的显著延迟和异常有关。与野生型形态菌株相比,ugmAΔ菌株具有异常的菌丝形态,产生宽、不均匀、高度分支的菌丝,通过透射电子显微镜观察可见其壁厚且相对电子密度高。在高渗培养基上或含有10μg/mL荧光增白剂的培养基上生长可部分修复这些影响,这与Galf在细胞壁结构和/或功能中起重要作用一致。