Late Embryogenesis Abundant (LEA) proteins confer water stress tolerance to mammalian somatic cells

Autor: L. Valbonetti, Antonella Fidanza, F.P. Luongo, Marta Czernik, P. A. Scapolo, Pasqualino Loi, Pasquale Patrizio
Rok vydání: 2020
Předmět:
Xeroprotectants
Somatic cell
030303 biophysics
Cell
Embryonic Development
Mitochondrion
Biology
Zea mays
General Biochemistry
Genetics and Molecular Biology
Desiccation tolerance
03 medical and health sciences
Cryoprotective Agents
0302 clinical medicine
medicine
Cold acclimation
Animals
Air drying
Late embryogenesis abundant (LEA) proteins
Mammalian somatic cells
Viability assay
Desiccation
Cryptobiosis
Cytoskeleton
Cells
Cultured
Triticum
Plant Proteins
030304 developmental biology
Cryopreservation
0303 health sciences
Sheep
030219 obstetrics & reproductive medicine
Dehydration
0402 animal and dairy science
Water
04 agricultural and veterinary sciences
General Medicine
Transfection
040201 dairy & animal science
Mitochondria
Cell biology
medicine.anatomical_structure
Artemia
General Agricultural and Biological Sciences
Zdroj: Cryobiology
ISSN: 0011-2240
DOI: 10.1016/j.cryobiol.2020.01.009
Popis: Late Embryogenesis Abundant (LEA) proteins are commonly found in plants and other organisms capable of undergoing severe and reversible dehydration, a phenomenon termed "anhydrobiosis". Here, we have produced a tagged version for three different LEA proteins: pTag-RAB17-GFP-N, Zea mays dehydrin-1dhn, expressed in the nucleo-cytoplasm; pTag-WCOR410-RFP, Tricum aestivum cold acclimation protein WCOR410, binds to cellular membranes, and pTag-LEA-BFP, Artemia franciscana LEA protein group 3 that targets the mitochondria. Sheep fibroblasts transfected with single or all three LEA proteins were subjected to air drying under controlled conditions. After rehydration, cell viability and functionality of the membrane/mitochondria were assessed. After 4 h of air drying, cells from the un-transfected control group were almost completely nonviable (1% cell alive), while cells expressing LEA proteins showed high viability (more than 30%), with the highest viability (58%) observed in fibroblasts expressing all three LEA proteins. Growth rate was markedly compromised in control cells, while LEA-expressing cells proliferated at a rate comparable to non-air-dried cells. Plasmalemma, cytoskeleton and mitochondria appeared unaffected in LEA-expressing cells, confirming the protection conferred by LEA proteins on these organelles during dehydration stress. This is likely to be an effective strategy when aiming to confer desiccation tolerance to mammalian cells.
Databáze: OpenAIRE
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