Photoconversion of matrix targeted GFP enables analysis of continuity and intermixing of the mitochondrial lumen

FEBS Letters

Hell, Stefan W.¹  Jakobs, Stefan¹  Schauss, Astrid C.¹ 

[1] Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany

关键词: Drp1;    Membrane fission;    Mitochondrial dynamics;    Photoactivation;    Photoinduction;    Saccharomyces cerevisiae;    GFP;    green fluorescent protein;   

DOI  :  10.1016/S0014-5793(03)01170-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We establish photoconversion of green fluorescent protein (GFP) as an optical ‘highlighter’ to investigate the continuity of the mitochondrial matrix in living budding yeast (Saccharomyces cerevisiae). Photoconversion of GFP resulting in a marked shift of the absorption and emission spectra to longer wavelengths is elicited, under low oxygen conditions, by irradiation with blue light. Photoconversion induced a several 100-fold increase in red fluorescence of matrix targeted GFP without affecting cell viability. The color changing facilitates simple and effective regional optical marking in a conventional fluorescence microscope. We found the mitochondrial compartment of S. cerevisiae to generally consist of one luminally continuous large part and occasionally some additional smaller fragments. Separated fragments fuse within a few minutes to the large part, resulting in a rapid intermixing of the entire mitochondrial matrix compartment. In Δfis1 and Δdnm1 mutants restricted in outer membrane fission, the mitochondria are still luminally continuous, suggesting a tight coupling of inner and outer membrane fissions. Matrix constrictions frequently occurring in wild type cells as well as in Δfis1 and Δdnm1 mutants do not interfere with luminal continuity.

【 授权许可】

Unknown