Xrn1 Exoribonuclease-An Intrinsic Marker of Yeast Population Growth.

Tomas Grousl, Tomas Vomastek
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Abstract

Background: Xrn1 exoribonuclease is the major mRNA degradation enzyme in Saccharomyces cerevisiae. In exponentially growing cells, Xrn1 is localised in the yeast cells and directs the degradation of mRNA molecules. Xrn1 is gradually deposited and presumably inactivated in the processing bodies (P-bodies) as the yeast population ages. Xrn1 can also localise to the membrane compartment of the arginine permease Can1/eisosome compartment at the yeast plasma membrane. This localisation correlates with the metabolic (diauxic) shift from glucose fermentation to respiration, although the relevance of this Xrn1 localisation remains unknown.

Methods: We monitored the growth rates and morphology of Xrn1-green fluorescent protein (GFP) cells compared to wild-type and Δxrn1 cells and observed the Xrn1-GFP localisation pattern in different media types for up to 72 hours using fluorescence microscopy.

Results: We present the dynamic changes in the localisation of Xrn1 as a versatile tool for monitoring the growth of yeast populations at the single-cell level using fluorescence microscopy.

Conclusions: The dynamic changes in the localisation of Xrn1 can be a versatile tool for monitoring the growth of yeast populations at the single-cell level. Simultaneously, Xrn1 localisation outside of P-bodies in post-diauxic cells supports its storage and cytoprotective function, yet the role of P-bodies in cell metabolism has still not yet been entirely elucidated.

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Xrn1 外核酸酶--酵母种群增长的内在标志物
背景:Xrn1 外切核酸酶是酿酒酵母(Saccharomyces cerevisiae)中主要的 mRNA 降解酶。在呈指数增长的细胞中,Xrn1 定位于酵母细胞内,指导 mRNA 分子的降解。随着酵母群体的老化,Xrn1 会逐渐沉积在加工体(P-bodies)中并可能失活。Xrn1 还能定位到酵母质膜上精氨酸渗透酶 Can1/eisosome 的膜区室。这种定位与从葡萄糖发酵到呼吸的代谢(双向)转变有关,但 Xrn1 定位的相关性仍不清楚:方法:与野生型细胞和Δxrn1细胞相比,我们监测了Xrn1-绿色荧光蛋白(GFP)细胞的生长速度和形态,并使用荧光显微镜观察了Xrn1-GFP在不同培养基类型中长达72小时的定位模式:我们将Xrn1定位的动态变化作为一种多功能工具,利用荧光显微镜在单细胞水平上监测酵母群体的生长情况:Xrn1定位的动态变化是在单细胞水平监测酵母种群生长的多功能工具。同时,Xrn1在后二叠体细胞中P体外的定位支持其储存和细胞保护功能,但P体在细胞新陈代谢中的作用尚未完全阐明。
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