Biochemical and structural characterization of DNA ligases from bacteria and archaea

DNA ligases are enzymes that seal breaks in the backbones of DNA, leading to them being essential for the survival of all organisms. DNA ligases have been studied from many different types of cells and organisms and shown to have diverse sizes and sequences, with well conserved specific sequences that are required for enzymatic activity. A significant number of DNA ligases have been isolated or prepared in recombinant forms and, here, we review their biochemical and structural characterization. All DNA ligases contain an essential lysine that transfers an adenylate group from a co-factor to the 5′-phosphate of the DNA end that will ultimately be joined to the 3′-hydroxyl of the neighbouring DNA strand. The essential DNA ligases in bacteria use β-nicotinamide adenine dinucleotide (β-NAD+) as their co-factor whereas those that are essential in other cells use adenosine-5′-triphosphate (ATP) as their co-factor. This observation suggests that the essential bacterial enzyme could be targeted by novel antibiotics and the complex molecular structure of β-NAD+ affords multiple opportunities for chemical modification. Several recent studies have synthesized novel derivatives and their biological activity against a range of DNA ligases has been evaluated as inhibitors for drug discovery and/or non-natural substrates for biochemical applications. Here, we review the recent advances that herald new opportunities to alter the biochemical activities of these important enzymes. The recent development of modified derivatives of nucleotides highlights that the continued combination of structural, biochemical and biophysical techniques will be useful in targeting these essential cellular enzymes.Several recent studies have synthesised novel derivatives and their biological activity against a range of DNA ligases has been evaluated as inhibitors for drug discovery and/or nonnatural substrates for biochemical applications. Here, we review the recent advances that herald new opportunities to alter the biochemical activities of these important enzymes. The recent development of modified derivatives of nucleotides highlights that the continued combination of structural, biochemical and biophysical techniques will be useful in targeting these essential cellular enzymes

Integration och validering av flygprogramvara för nanosatelliter inom projektet ESA OPS-SAT

With the increasing number of satellites operating in orbit and the development of nanosatelliteconstellations, it has become more and more arduous for operators to keep track of every satellitestate, and perform corrective or avoidance manoeuvres. That is why CNES, the French space agency,is developing new algorithms, which aimed at making satellites more self-su cient. More especially,these algorithms are in charge of autonomous orbit control, collision risk calculations and satellitestatus monitoring. In this thesis, we present the architecture of these three algorithms and how theyinteract between them to deal with the autonomous control of a satellite. In addition, this paper studiestheir integration within the OPS-SAT nanosatellite, which is an in-orbit demonstrator developed bythe European Space Agency (ESA) and opened to worldwide experimenters. By analysing the dataused by the numerical propagators, the size of the input configuration files sent to the nanosatellitewas optimised. Thanks to this optimisation, the size of telecommands sent during each OPS-SATflyby above the ESOC ground station meets the requirements. Due to some issues encountered with the nanosatellite’s GPS, a solution was found to update thecurrent orbit on-board, and thus allow the proper algorithms’ operation. This thesis also introduceshow the tests were carried out in order to validate these algorithms, both on flat-sat and on the realsatellite. The results demonstrate that their integration on the OPS-SAT numerical environment issuccessful, meaning that the algorithms and their dependences are correctly packaged, sent and uploaded,and that they work as expected. Their execution time are of course longer due to the limitedcalculation capacity of the on-board computer, but are still compatible with real operations, except forthe collision risk computation, which can exceed the orbital period depending on the initial conditions.Finally, the thesis presents the process of real operations for one of the three algorithms developed byCNES, the di culties encountered and the solutions considered.Med det ökande antalet satelliter i omloppsbana och utvecklingen av nanosatellitkonstellationer hardet blivit mer och mer krävande för operatörer att hålla reda på varje satellits tillstånd och utförakorrigerande eller undvikande manövrar. Det är därför som CNES, den franska rymdorganisationen,utvecklar nya algoritmer som syftar till att göra satelliter mer autonoma. Närmare bestämt ansvarardessa algoritmer för autonom omloppsbanereglering, kollisionsriskberäkningar och satellitstatusövervakning.I detta examensarbete presenterar vi arkitekturen för dessa tre algoritmer och hur de interagerarmellan sig för att hantera den autonoma styrningen av en satellit. Dessutom studeras deras integrationinom OPS-SAT-nanosatelliten, som är en demonstrator i omloppsbana som utvecklats av Europeiskarymdorganisationen (ESA) och öppnad för globala experiment. Genom att analysera de datasom används av de numeriska propagatorerna optimerades storleken på de ingångskonfigurationsfilersom skickades till nanosatelliten. Tack vare denna optimering uppfylls storlekskraven på telekommandonsom skickas under varje passage av OPS-SAT ovanför ESOC-markstationen. På grund av vissa problem med nanosatellitens GPS hittades en lösning för att uppdatera den aktuellaomloppsbanan ombord och därmed möjliggöra korrekt funktion av algoritmerna. Detta examensarbeteintroducerar också hur testerna genomfördes för att validera dessa algoritmer, både på en s.k. flat-satoch på den verkliga satelliten. Resultaten visar att deras integration i den numeriska miljön OPS-SATär framgångsrik, vilket innebär att algoritmerna och deras beroende är korrekt förpackade, skickade ochuppladdade och att de fungerar som förväntat. Deras exekveringstid är naturligtvis längre på grundav den inbyggda datorns begränsade beräkningskapacitet, men är fortfarande kompatibel med verkligaoperationer, förutom beräkningen av kollisionsrisk, som kan överstiga omloppsperioden beroende påde initiala förhållandena. Slutligen presenterar rapporten processen för verkliga operationer för en avde tre algoritmerna som utvecklats av CNES, svårigheterna och de lösningar som övervägs