Thermoprotection by a cell membrane-localized metacaspase in a green alga

Caspases are restricted to animals, while other organisms, including plants, possess metacaspases (MCAs), a more ancient and broader class of structurally related yet biochemically distinct proteases. Our current understanding of plant MCAs is derived from studies in streptophytes, and mostly in Arabidopsis (Arabidopsis thaliana) with 9 MCAs with partially redundant activities. In contrast to streptophytes, most chlorophytes contain only 1 or 2 uncharacterized MCAs, providing an excellent platform for MCA research. Here we investigated CrMCA-II, the single type-II MCA from the model chlorophyte Chlamydomonas (Chlamydomonas reinhardtii). Surprisingly, unlike other studied MCAs and similar to caspases, CrMCA-II dimerizes both in vitro and in vivo. Furthermore, activation of CrMCA-II in vivo correlated with its dimerization. Most of CrMCA-II in the cell was present as a proenzyme (zymogen) attached to the plasma membrane (PM). Deletion of CrMCA-II by genome editing compromised thermotolerance, leading to increased cell death under heat stress. Adding back either wild-type or catalytically dead CrMCA-II restored thermoprotection, suggesting that its proteolytic activity is dispensable for this effect. Finally, we connected the non-proteolytic role of CrMCA-II in thermotolerance to the ability to modulate PM fluidity. Our study reveals an ancient, MCA-dependent thermotolerance mechanism retained by Chlamydomonas and probably lost during the evolution of multicellularity. A plasma membrane-localized metacaspase mediates membrane fluidity and confers thermotolerance in Chlamydomonas reinhardtii independently of its proteolytic activity.

Automated summary

Plants possess a more ancient and broader class of proteases called MetaCaspases (MCAs) in contrast to the animal-specific Caspases. This study investigates a single type-II MCA called CrMCA-II from the model chlorophyte Chlamydomonas reinhardtii. Surprisingly, CrMCA-II dimerizes like Caspases, and its activation is correlated with dimerization. Most of CrMCA-II in the cell is in the form of a proenzyme attached to the plasma membrane. Deletion of CrMCA-II compromises thermotolerance, but adding back either wild-type or catalytically dead CrMCA-II restores thermoprotection, suggesting that its proteolytic activity is not essential for its thermotolerance effect. Furthermore, CrMCA-II's non-proteolytic role is found to be related to the modulation of plasma membrane fluidity.