Project description:The present study describes the isolation of a Thermococcus sp. strain 175 from the world‘s deepest hydrothermal vent sites known thus far – The Mid-Cayman Rise.consisting of two hydrothermal venting systems Bee Bee (or Piccard), at 4950m depth and Von Damm (or Walsh) at 2300m The strain is capable of growth over 0.1MPa (atm. Pressure) to 120MPa, the widest known range of pressure dependent growth. The study further explores piezophilic adaptation using comparative genomic tools. Insights into the transcriptome of this strain providers the first look into the transcriptional machinery of peizophilic Thermococci.
Project description:Bathymodiolus azoricus is a deep-sea mussel found in the hydrothermal vent fields of the Mid-Atlantic Ridge. It lives in symbiosis with sulfur- and methane-oxidizing γ-proteobacteria within its gills. In our study, we aimed to understand the metabolic and physiological interconnections between the symbiotic partners. For this purpose, symbionts and host were physically separated using density gradient centrifugation. This procedure yielded a symbiont-enriched gradient pellet fraction and a supernatant fraction enriched in host components. The cytosolic and membrane-associated proteome of both these fractions along with whole gill and foot tissue of the mussel were then investigated through 1D-PAGE LC-MS/MS. Proteins were quantified based on their spectral counts using the NSAF method. For efficient identification, sequences from evolutionarily related endosymbiotic and free-living bacteria and from bivalve host relatives were compiled into a comprehensive protein database. A total of 3178 host and symbiont proteins were identified from all samples.