Temporal profiling of redox-dependent heterogeneity in single cells

eLife

Sidra Ilyas¹  Dana Reichmann¹  Sabina Tsytkin-Kirschenzweig²  Yifat Geffen²  Meytal Radzinski²  Ohad Yogev²  Nir Friedman²  Rosi Fassler²  Jenia Gutin²  Maya Schuldiner²  Nadav Shai³  Tommer Ravid⁴  Yaakov Nahmias⁴  William Breuer⁵ 

[1] School of Computer Science and Engineering, The Hebrew University of Jerusalem, Jerusalem, Israel;Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, Safra Campus Givat Ram, The Hebrew University of Jerusalem, Jerusalem, Israel;Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel;Grass Center for Bioengineering, Benin School of Computer Science and Engineering, The Hebrew University of Jerusalem, Jerusalem, Israel;Proteomics and Mass Spectrometry Unit, The Alexander Silberman Institute of Life Sciences, Safra Campus Givat Ram, The Hebrew University of Jerusalem, Jerusalem, Israel;

关键词: redox sensors;    redox biology;    yeast;    roGFP;   

DOI  :  10.7554/eLife.37623

来源: DOAJ

【 摘 要 】

Cellular redox status affects diverse cellular functions, including proliferation, protein homeostasis, and aging. Thus, individual differences in redox status can give rise to distinct sub-populations even among cells with identical genetic backgrounds. Here, we have created a novel methodology to track redox status at single cell resolution using the redox-sensitive probe Grx1-roGFP2. Our method allows identification and sorting of sub-populations with different oxidation levels in either the cytosol, mitochondria or peroxisomes. Using this approach, we defined a redox-dependent heterogeneity of yeast cells and characterized growth, as well as proteomic and transcriptomic profiles of distinctive redox subpopulations. We report that, starting in late logarithmic growth, cells of the same age have a bi-modal distribution of oxidation status. A comparative proteomic analysis between these populations identified three key proteins, Hsp30, Dhh1, and Pnc1, which affect basal oxidation levels and may serve as first line of defense proteins in redox homeostasis.

【 授权许可】

Unknown