Hurtado Cleofe A R, Rachubinski Richard A
Department of Cell Biology, University of Alberta, Medical Sciences Building 5-14, Edmonton, Alberta T6G 2H7, Canada1.
Microbiology (Reading). 2002 Nov;148(Pt 11):3725-3735. doi: 10.1099/00221287-148-11-3725.
Most pathogenic fungi have the ability to alternate between a unicellular yeast form and different filamentous forms (hyphae and pseudohyphae). This attribute is generally regarded as an important virulence factor and has also attracted attention because of its implications in the study of eukaryotic cell differentiation. To identify genes that are involved in the regulation of these events, chemical mutagenesis of the dimorphic yeast Yarrowia lipolytica was performed and morphological mutants that were unable to form hyphal cells were isolated. Screening of a Y. lipolytica genomic DNA library for genes able to complement this defect led to the isolation of YlBMH1, a gene encoding a 14-3-3 protein and whose transcription levels are increased during the yeast-to-hypha transition. Remarkably, overexpression of YlBMH1 was able to enhance pseudohyphae formation in a strain lacking functional YlRAC1 but caused no visible effects in deltamhy1 and deltabem1 cells, thus suggesting that YlBMH1 is involved in the regulation of both hyphal and pseudohyphal growth in Y. lipolytica. The identification of YlBMH2, a gene encoding a second 14-3-3 protein (YlBmh2p) that contains a 19 aa insertion absent in all other members of the 14-3-3 family, is also reported. Differently from YlBMH1, the transcription levels of YlBMH2 do not show any apparent variation during the induction of hyphal growth, and its overexpression has no effects on cells lacking functional MHY1, YlRAC1 or YlBEM1. Taken together, these observations suggest that, in spite of their high conservation, YlBmh1p and YlBmh2p have different cellular functions.
大多数致病真菌能够在单细胞酵母形态和不同的丝状形态(菌丝和假菌丝)之间转换。这一特性通常被视为一种重要的毒力因子,并且因其在真核细胞分化研究中的意义也受到了关注。为了鉴定参与调控这些过程的基因,对二态酵母解脂耶氏酵母进行了化学诱变,并分离出了无法形成菌丝细胞的形态突变体。通过筛选解脂耶氏酵母基因组DNA文库中能够弥补这一缺陷的基因,分离出了YlBMH1,该基因编码一种14-3-3蛋白,其转录水平在酵母向菌丝转变过程中升高。值得注意的是,YlBMH1的过表达能够增强缺乏功能性YlRAC1的菌株中的假菌丝形成,但在deltamhy1和deltabem1细胞中没有产生明显影响,因此表明YlBMH1参与了解脂耶氏酵母中菌丝和假菌丝生长的调控。本文还报道了YlBMH2的鉴定,该基因编码第二个14-3-3蛋白(YlBmh2p),在14-3-3家族的所有其他成员中都不存在一个19个氨基酸的插入序列。与YlBMH1不同,YlBMH2的转录水平在菌丝生长诱导过程中没有显示出任何明显变化,并且其过表达对缺乏功能性MHY1、YlRAC1或YlBEM1的细胞没有影响。综上所述,这些观察结果表明,尽管YlBmh1p和YlBmh2p具有高度保守性,但它们具有不同的细胞功能。