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Xrn1 Exoribonuclease-An Intrinsic Marker of Yeast Population Growth
Tomas Grousl1, Tomas Vomastek1
1Laboratory of Cell Signalling, Institute of Microbiology of the Czech Academy of Sciences, 14200 Prague, Czech Republic.
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.
Insights
The exoribonuclease Xrn1
Area of Science:
- Cell biology
- Molecular biology
- Yeast genetics
Background:
- Xrn1 exoribonuclease is the primary mRNA degradation enzyme in Saccharomyces cerevisiae.
- Xrn1 localization shifts from general cellular distribution in growing cells to processing bodies (P-bodies) in aging yeast populations.
- Xrn1 also localizes to the plasma membrane during the diauxic shift, a phenomenon with unknown significance.
Purpose of the Study:
- To investigate the dynamic localization of Xrn1 in yeast populations.
- To assess the utility of Xrn1 localization as an indicator of yeast growth and metabolic state.
- To explore the functional implications of Xrn1 localization outside of P-bodies.
Main Methods:
- Monitoring growth rates and morphology of Xrn1-green fluorescent protein (GFP) expressing cells.
- Comparing Xrn1-GFP localization patterns in different media over 72 hours.
- Utilizing fluorescence microscopy to observe dynamic Xrn1 localization.
Main Results:
- Dynamic changes in Xrn1 localization were observed.
- Xrn1 localization patterns were found to be a versatile indicator for monitoring yeast population growth at the single-cell level.
- Localization of Xrn1 outside P-bodies in post-diauxic cells was confirmed.
Conclusions:
- Dynamic Xrn1 localization serves as a valuable tool for single-cell level monitoring of yeast population growth.
- Xrn1 localization outside P-bodies suggests a storage and cytoprotective role in post-diauxic cells.
- The precise role of P-bodies in yeast cell metabolism requires further elucidation.
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