Laboratory of Cell Signalling, Institute of Microbiology of the Czech Academy of Sciences, 14200 Prague, Czech Republic.
Front Biosci (Elite Ed). 2024 Jan 31;16(1):1. doi: 10.31083/j.fbe1601001.
Xrn1 exoribonuclease is the major mRNA degradation enzyme in 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.
We monitored the growth rates and morphology of Xrn1-green fluorescent protein (GFP) cells compared to wild-type and Δ cells and observed the Xrn1-GFP localisation pattern in different media types for up to 72 hours using fluorescence microscopy.
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.
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.
Xrn1 外切核酸酶是酵母细胞中主要的 mRNA 降解酶，在指数生长期的细胞中，Xrn1 定位于酵母细胞中，并指导 mRNA 分子的降解。随着酵母群体的老化，Xrn1 逐渐沉积并可能失活于处理体（P 体）中。Xrn1 也可以定位到酵母质膜上的精氨酸渗透酶 Can1/核内体隔室的膜隔室。这种定位与从葡萄糖发酵到呼吸的代谢（兼性）转变相关联，尽管这种 Xrn1 定位的相关性仍然未知。
我们监测了 Xrn1-绿色荧光蛋白（GFP）细胞与野生型和Δ细胞的生长速度和形态，并在不同的培养基类型中使用荧光显微镜观察了长达 72 小时的 Xrn1-GFP 定位模式。
我们展示了 Xrn1 定位的动态变化，这是一种使用荧光显微镜在单细胞水平监测酵母群体生长的多功能工具。
Xrn1 定位的动态变化可以成为一种监测单细胞水平酵母群体生长的多功能工具。同时，在兼性细胞中 P 体外的 Xrn1 定位支持其储存和细胞保护功能，但 P 体在细胞代谢中的作用仍未完全阐明。
