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Engineering yeast endosymbionts as a first step towards laboratory evolution of mitochondria
Author(s) -
Mehta Angad,
Supekova Lubica,
Supek Frantisek,
Schultz Peter
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.530.3
Subject(s) - endosymbiosis , biology , yeast , saccharomyces cerevisiae , auxotrophy , escherichia coli , mitochondrion , bacteria , intracellular parasite , biochemistry , intracellular , microbiology and biotechnology , gene , genetics , chloroplast , plastid
It has been hypothesized that mitochondria evolved from a bacterial ancestor that initially became established in a protoeukaryotic cell as an endosymbiont. Here we model this first stage of mitochondrial evolution by engineering endosymbiosis between E. coli and the budding yeast S. cerevisiae . Fusion of yeast with E. coli ectopically expressing several genes from unrelated, intracellular bacteria was key for establishing endosymbiosis. ADP/ATP translocase‐expressing E. coli provided an energy source for a respiration‐deficient cox2 yeast mutant, enabling growth of yeast‐ E. coli chimera on a non‐fermentable carbon source. Similarly, yeast provided a source of thiamin or NAD to an E. coli thiamin or NAD auxotroph respectively. Expression of several SNARE‐like protein on the surface of E. coli was also required to prevent lysosomal degradation. The engineered yeast‐ E. coli chimeras sustained growth on selection medium in containing the antibiotic carbenicillin indicating the presence of intracellular E. coli which supports the growth by ATP synthesis on selection medium. Further, sf‐gfp expressing E. coli endosymbionts could be observed in the yeast by super resolution fluorescence microscopy after more than 40 doublings. This readily manipulated system should allow us to experimentally delineate host‐endosymbiont adaptations that occurred during evolution of the current much reduced mitochondrial genome This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .