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Updated: Jun 7, 2025

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Published on: October 22, 2018

Mapping Active RNA Polymerases in Proliferating and Quiescent Fission Yeast Cells Using Precision Run-On Sequencing.

Alicia Vázquez-Bolado¹, Pei-Yun Jenny Wu²

¹Institute of Biochemistry and Cellular Genetics, CNRS UMR 5095 and University of Bordeaux, Bordeaux, France.

Methods in Molecular Biology (Clifton, N.J.)

|November 11, 2024

Summary

We optimized the quantitative Precision nuclear Run-On sequencing (qPRO-seq) method for fission yeast. This technique maps RNA polymerase activity genome-wide, even under nutrient-limiting conditions.

Keywords:

Nascent RNA Nitrogen starvation PRO-seq Quiescence S. pombe Transcription

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Published on: March 7, 2019

Area of Science:

Molecular Biology
Genomics
Transcriptional Regulation

Background:

Next-generation sequencing (NGS) has advanced the study of transcriptional regulation.
Precision nuclear Run-On sequencing (PRO-seq) maps RNA polymerase active sites genome-wide.
Initial PRO-seq protocols were sample-intensive and technically challenging.

Purpose of the Study:

To adapt and validate the quantitative PRO-seq (qPRO-seq) method for fission yeast (Schizosaccharomyces pombe).
To assess RNA polymerase activity in fission yeast under nutrient-limiting conditions.

Main Methods:

Adaptation and optimization of the qPRO-seq protocol for Schizosaccharomyces pombe.
Application of qPRO-seq to analyze transcriptional activity in proliferating and quiescent yeast cells under nutrient limitation.

Main Results:

Successfully validated and optimized qPRO-seq for fission yeast.
Demonstrated the utility of qPRO-seq for studying RNA polymerase activity in nutrient-stressed yeast.

Conclusions:

The optimized qPRO-seq protocol provides a robust method for high-resolution analysis of transcriptional activity in fission yeast.
This technique is valuable for investigating gene regulation in response to environmental cues, such as nutrient availability.