The dynamin-like protein Fzl promotes thylakoid fusion and resistance to light stress in Chlamydomonas reinhardtii

Large GTPases of the Dynamin Related Proteins (DRP) family shape lipid bilayers through membrane fission or fusion processes. Despite the highly organized photosynthetic membranes of thylakoids, a single DRP is known to be targeted inside the chloroplast. Fzl from the land plant Arabidopsis thaliana is inserted in the inner envelope and thylakoid membranes to regulate their morphology. Fzl may promote the fusion of thylakoids but this remains to be proven. Moreover, the physiological requirement for fusing thylakoids is currently unknown. Here, we find that the unicellular microalga Chlamydomonas reinhardtii encodes an Fzl ortholog (CrFzl) that is localized in the chloroplast where it is soluble. To explore its function, the CRISPR/Cas9 technology was employed to generate multiple CrFzl knock out strains. Phenotypic analyzes revealed a specific requirement of CrFzl for survival upon light stress. Consistent with this, strong irradiance lead to increased photoinhibition of photosynthesis in mutant cells. Fluorescence and electron microscopy analysis demonstrated that upon exposure to high light, CrFzl mutants show defects in chloroplast morphology but also large cytosolic vacuoles in close contact with the plastid. We further observe that strong irradiance induces an increased recruitment of the DRP to thylakoid membranes. Most importantly, we show that CrFzl is required for the fusion of thylakoids during mating. Together, our results suggest that thylakoids fusion may be necessary for resistance to light stress. Author summary All eukaryotic cells are composed of compartments with defined functions. Among those, mitochondria generate the main source of energy in human and animal cells. Their capacity to generate and diffuse energy in the cell is regulated by fusion and fragmentation processes. Together with mitochondria that produce energy from oxygen, plant cells include an additional compartment called the chloroplast that produces energy from light. The machinery that converts light into energy is more precisely located inside the chloroplast within stacks of membranes called the thylakoids. Here, we elucidate the function of CrFzl, a previously uncharacterized protein encoded by the genome of the unicellular alga Chlamydomonas reinhardtii. Algal cells that do not contain CrFzl are impaired in their capacity to grow when they receive too much light and our results indicate that CrFzl promotes the fusion of thylakoids during mating. These results suggest that membrane fusion is an essential tool for energy production in stress conditions by living organisms.