G-Equivalent Acceleration Tolerance in the Eutardigrade Species Hypsibius dujardini.

Tardigrades are microscopic organisms renowned for their ability to survive extreme environmental conditions. Tardigrade extreme-tolerance research has centered on the ability to withstand desiccation, low and high temperatures, and high hydrostatic pressure and radiation levels. Tardigrade tolerance to hypergravity, however, has yet to be described. We used the eutardigrade species Hypsibius dujardini to investigate short-term tolerance to g-equivalent accelerations (i.e., mimicking g-forces). Data obtained from specimens centrifuged between 3421g and 16,060g for 1 min inclusively reveal tolerance in an acceleration-dependent relation, with lower survivorship and egg production at higher accelerations. This is the first study to demonstrate tardigrade potential for tolerance to hypergravity and describe expected effects on tardigrade survival and reproduction. These findings will prove to be useful in lithopanspermia research (i.e., viable spread in meteoritic rocks). Key Words: Astrobiology-Extreme tolerance-Hypergravity-Tardigrade. Astrobiology 17, 55-60.

Structural determinants stabilizing the axial channel of ClpP for substrate translocation.

Acyldepsipeptides (ADEPs) antibiotics bind to Escherichia coli ClpP mimicking the interactions that the IGL/F loops in ClpA or ClpX ATPases establish with the hydrophobic pockets surrounding the axial pore of the tetradecamer that the protease forms. ADEP binding induces opening of the gates blocking the axial channel of ClpP and allowing protein substrates to be translocated and hydrolysed in the degradation chamber. To identify the structural determinants stabilizing the open conformation of the axial channel for efficient substrate translocation, we constructed ClpP variants with amino acid substitutions in the N-terminal region that forms the axial gates. We found that adoption of a ß-hairpin loop by this region and the integrity of the hydrophobic cluster at the base of this loop are necessary elements for the axial gate to efficiently translocate protein substrates. Analysis of ClpP variants from Bacillus subtilis suggested that the identified structural requirements of the axial channel for efficient translocation are conserved between Gram-positive and Gram-negative bacteria. These findings provide mechanistic insights into the activation of ClpP by ADEPs as well as the gating mechanism of the protease in the context of the ClpAP and ClpXP complexes.