Meeting Abstract
Adaptations in animals to survive severe desiccation (anhydrobiosis) are multifaceted and include expression of highly hydrophilic polypeptides termed late embryogenesis abundant (LEA) proteins. Several classification schemes for LEA proteins have been proposed. However, the brine shrimp, Artemia franciscana, is the only known animal that naturally expresses LEA proteins from three different classification groups (groups 1, 3, and 6). We hypothesized that proteins from groups 1 and 3 may function to aid cells in entering the anhydrobiotic state, while group 6 LEA proteins may be required to prolong viability in the dry state, or to ameliorate cellular damage during rehydration. To test our hypothesis, cell lines that transgenically express combinations of different LEA proteins have to be developed. We utilized Kc167 cells from the desiccation sensitive organism Drosophila melanogaster to express LEA proteins belonging to groups 3 and 6 concurrently from the same multicistronic vector construct by employing viral derived cis-acting hydrolase element peptides. Proteins encoded on the same mRNA strand are separated during translation in a process described as ‘ribosome-skipping’. Despite bacterial recombination of vector constructs during cloning of plasmids, our protein immunoblots illustrated that concurrent expression of multiple LEA proteins is possible in Kc167 cells. Experiments to investigate the effect of different combinations of LEA proteins belonging to group 3 and 6 on viability of Kc167 cells after rapid desiccation and rehydration are currently underway. Supported by NSF IOS-1457061/IOS-1456809.
