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Proteinaceous liquid-liquid phase separation (LLPS) occurs when a polypeptide coalesces into a dense phase to form a liquid droplet (i.e., condensate) in aqueous solution. In vivo, functional protein-based condensates are often referred to as membraneless organelles (MLOs), which have roles in cellular processes ranging from stress responses to regulation of gene expression. Late embryogenesis abundant (LEA) proteins containing seed maturation protein domains (SMP; PF04927) have been linked to storage tolerance of orthodox seeds. The mechanism by which anhydrobiotic longevity is improved is unknown. Interestingly, the brine shrimp
Artemia franciscana is the only animal known to express such a protein (Afr LEA6) in its anhydrobiotic embryos. Ectopic expression ofAfr LEA6 (AWM11684) in insect cells improves their desiccation tolerance and a fraction of the protein is sequestered into MLOs, while aqueousAfr LEA6 raises the viscosity of the cytoplasm. LLPS ofAfr LEA6 is driven by the SMP domain, while the size of formed MLOs is regulated by a domain predicted to engage in protein binding.Afr LEA6 condensates formed in vitro selectively incorporate target proteins based on their surface charge, while cytoplasmic MLOs formed inAfr LEA6-transfected insect cells behave like stress granules. We suggest thatAfr LEA6 promotes desiccation tolerance by engaging in two distinct molecular mechanisms: by raising cytoplasmic viscosity at even modestmore »