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The deep ocean holds a diversity of life that remains to be discovered and described. Marine ecosystems fueled by chemosynthesis, such as hydrothermal vents and hydrocarbon seeps, represent biodiversity hotspots along the ocean floor. Yet, investigations into the diversity, genetic connectivity, and biogeography of life at such ecosystems are still ongoing. The present dissertation explores the diversity and biogeography of deep-sea gastropod mollusks from hydrocarbon seeps at the Pacific Costa Rica Margin to understand what factors lead to the generation and maintenance of novel biodiversity at chemosynthesis-based ecosystems in the ocean. The Pacific Costa Rica Margin was extensively sampled and explored between 2017 and 2019 and yielded an abundance and diversity of marine life. Gastropod mollusks were chosen as the model organisms for this research as they are taxonomically diverse, are found in high abundance, and occur at chemosynthesis-based ecosystems worldwide. Integrating taxonomic, phylogenetic, population genetic, and biogeographic methods, this dissertation contributes new species, occurrence records, genetic data, taxonomic tools, and biogeographic knowledge for deep-sea gastropods. As global biodiversity continues to be threatened by anthropogenic impacts such as overexploitation and climate change, documenting and understanding deep-sea biodiversity and the factors that influence it is of critical importance. / Biology

Chemosynthetic ecosystems are habitats whose food webs rely on chemosynthesis, a process by which bacteria fix carbon using energy from chemicals, rather than sunlight-driven photosynthesis for primary production, and they are found all over the world on the ocean floor. Although these deep-sea habitats are remote, they are increasingly being impacted by human activities such as oil and gas exploration and the imminent threat of deep-sea mining. My dissertation examines deep-sea chemosynthetic ecosystems at several ecological scales to answer basic biology questions and lay a foundation for future researchers studying these habitats. There are two major varieties of chemosynthetic ecosystems, hydrothermal vents and cold seeps, and my dissertation studies both. My first chapter begins at cold seeps and at the population level by modeling the population dynamics and lifespan of a single species of tubeworm, Escarpia laminata, found in the Gulf of Mexico. I found that this tubeworm, a foundation species that forms biogenic habitat for other seep animals, can reach ages over 300 years old, making it one of the longest-lived animals known to science. According to longevity theory, its extreme lifespan is made possible by the stable seep environment and lack of extrinsic mortality threats such as predation. My second chapter expands the scope of my research from this single species to the entire cold seep community and surrounding deep-sea animals common to the Gulf of Mexico. The chemicals released at cold seeps are necessary for chemosynthesis but toxic to non-adapted species such as cold-water corals. Community studies in this area have previously shown that seeps shape community assembly through niche processes. Using fine-scale water chemistry samples and photographic mapping of the seafloor, I found that depressed dissolved oxygen levels and the presence of hydrogen sulfide from seepage affect foundation taxa distributions, but the concentrations of hydrocarbons released from these seeps did not predict the distributions of corals or seep species. In my third chapter I examine seep community assembly drivers in the Costa Rica Margin and compare the macrofaunal composition at the family level to both hydrothermal vents and methane seeps around the world. The Costa Rica seep communities have not previously been described, and I found that depth was the primary driver behind community composition in this region. Although this margin is also home to a hybrid “hydrothermal seep” feature, this localized habitat did not have any discernible influence on the community samples analyzed. When vent and seep communities worldwide were compared at the family-level, geographic region was the greatest determinant of community similarity, accounting for more variation than depth and habitat type. Hydrothermal vent and methane seeps are two chemosynthetic ecosystems are created through completely different geological processes, leading to extremely different habitat conditions and distinct sets of related species. However, at the broadest spatial scale and family-level taxonomic resolution, neutral processes and dispersal limitation are the primary drivers behind community structure, moreso than whether the habitat is a seep or a vent. At more local spatial scales, the abiotic environment of seeps still has a significant influence on the ecology of deep-sea organisms. The millennial scale persistence of seeps in the Gulf of Mexico shapes the life history of vestimentiferan tubeworms, and the sulfide and oxygen concentrations at those seeps determine seep and non-seep species’ distributions across the deep seafloor. / Biology

Peptide und Proteine sind wichtige Untersuchungsobjekte der biochemischen Forschung. Es wurden in den letzten Jahren eine Reihe von Ligationsmethoden entwickelt, um weitgehend entschützte, gereinigte Peptidsequenzen im wässrigen Milieu zu koppeln. Vor diesem Hintergrund von besonderem Interesse für einen möglichen Einsatz bei Ligationen ist die bakterielle Transpeptidase Sortase A. Dieses Enzym ist in vivo an der Anknüpfung von Proteinen an das bakterielle Peptidoglycan beteiligt, wobei es Substrate an einem LPXTG-Motiv zwischen Threonin und Glycin spaltet und auf ein Oligoglycin-Nucleophil überträgt. Zur Untersuchung der Enzymaktivität wurde in dieser Arbeit ein einfacher HPLC-basierter Assay etabliert. Die an Peptidmodellen gewonnenen Resultate wurden schließlich für den Aufbau eines löslichen Rezeptors genutzt. Ein Schlüsselschritt war die Sortase-vermittelte Ligation des in E. coli exprimierten, gefalteten Rezeptor-N-Terminus an ein 3-Loop-Konstrukt. Das erhaltene 23 kDa große Rezeptormimetikum war nach chromatographischer Reinigung homogen gemäß HPLC und MS. Es zeigte eine spezifische, hoch affine Bindung zu natürlichen Peptidliganden des CRF1-Rezeptors. Weiterhin konnte demonstriert werden, dass sich Sortase für die selektive Markierung von Proteinen eignet. So wurde ein Fluoreszenzlabel C-terminal an das 50 kDa Protein NEMO geknüpft. Als weiteres Anwendungsbeispiel der Sortase-vermittelten Ligation diente die Darstellung von PNA–CPP-Konjugaten. Die Verwendung von Überschüssen des Peptides und die Entfernung der niedermolekularen Abgangsgruppe durch Dialyse erwies sich als sehr effektiv und gestattete gute bis hervorragende Kupplungsausbeuten von bis zu 94%. Die biologische Wirkung der erhaltenen CPP–PNA-Konjugate konnte in Aufnahmeuntersuchungen an Zellen gezeigt werden. / Peptides and proteins are important research objects in biochemical research. Therefore, several ligation methods to couple unprotected, purified peptide sequences in aqueous media have been developed during the last years. At a special interest in this case is the bacterial transpeptidase sortase A. This enzyme couples proteins in vivo to the bacterial peptidoglycan by cleavage at a LPXTG-recognition motif between threonine and glycine and subsequent transfer to an oligoglycin nucleophile. In order to investigate the enzymatic activity, a simple HPLC-based assay was established in this work. Results obtained with model peptides were used for the assembly of a soluble receptor. A key step was the sortase-mediated ligation of the folded receptor N-terminus (expressed in E. coli) to the 3-loop-construct. The resulting receptor mimic of 23 kDa was homogeneous according to HPLC and MS. It showed specific binding to natural peptide ligands of the CRF1-receptor with high affinity. Furthermore, it could be shown that sortase is usable for selective protein labeling. For this purpose, a fluorescence label was attached C-terminally to the 50 kDa protein NEMO. As a further example of sortase-mediated ligation served the synthesis of PNA-CPP-conjugates. The use of an excess of the peptide and dialyzing away the small leaving group proved to be very effective and coupling yields up to 94% could be achieved. The biological activity of the CPP-PNA-conjugates could be shown by uptake studies in cells.