A Yeast Chromatin-enriched Fractions Purification Approach, yChEFs, from Saccharomyces cerevisiae

Abel Cuevas-Bermúdez; Ana I Garrido-Godino; Francisco Gutiérrez-Santiago; Francisco Navarro

doi:10.21769/BioProtoc.3471

A Yeast Chromatin-enriched Fractions Purification Approach, yChEFs, from Saccharomyces cerevisiae

Bio Protoc. 2020 Jan 5;10(1):e3471. doi: 10.21769/BioProtoc.3471.

Authors

Abel Cuevas-Bermúdez¹, Ana I Garrido-Godino¹, Francisco Gutiérrez-Santiago¹, Francisco Navarro^{1

2}

Affiliations

¹ Departamento de Biología Experimental-Genética, Universidad de Jaén, Jaén, Spain.
² Centro de Estudios Avanzados en Aceite de Oliva y Olivar. Universidad de Jaén, Paraje de las Lagunillas, s/n, E23071, Jaén, Spain.

Abstract

We have adapted a previous procedure and improved an approach that we named yChEFs (yeast Chromatin Enriched Fractions) for purifying chromatin fractions. This methodology allows the easy, reproducible and scalable recovery of proteins associated with chromatin. By using yChEFs, we bypass subcellular fractionation requirements involved when using zymolyase to obtain the spheroplast, which is employed in many other procedures. Employing small amount of culture cells and small volumes of solutions during the yChEFs procedure is very useful to allow many samples to be handled at the same time, and also reduces costs and efforts. The purified proteins associated with chromatin fractions obtained by yChEFs can be analyzed by Western blot (Figure 1) or combined with mass spectrometry for proteomic analyses.

Keywords: Chromatin; Mass spectrometry; Proteins; Saccharomyces cerevisiae; Yeast.