Spectroscopic characterization of exoplanets : from LOUPE to SINFONI

Over the past years it has been discovered that the population of extra-solar planets is large and diverse. This fact feeds expectations for finding habitable Earth-like planets and potentially extra-terrestrial life. However without a reliable characterization, the fundamental nature of these planets would remain unknown after their initial discovery. This thesis focuses on the development of new spectroscopic observation and analysis methods to characterize planets after their discovery. The techniques presented in this thesis can be used in the future to detect the signs of life and habitability on Earth-like exoplanets that are expected to be discovered over the coming years. Stars and planetary system

Deletion of the Nucleotide Excision Repair Gene Ercc1 Reduces Immunoglobulin Class Switching and Alters Mutations Near Switch Recombination Junctions

The structure-specific endonuclease ERCC1-XPF is an essential component of the nucleotide excision DNA repair pathway. ERCC1-XPF nicks double-stranded DNA immediately adjacent to 3′ single-strand regions. Substrates include DNA bubbles and flaps. Furthermore, ERCC1 interacts with Msh2, a mismatch repair (MMR) protein involved in class switch recombination (CSR). Therefore, ERCC1-XPF has abilities that might be useful for antibody CSR. We tested whether ERCC1 is involved in CSR and found that Ercc1−/− splenic B cells show moderately reduced CSR in vitro, demonstrating that ERCC1-XPF participates in, but is not required for, CSR. To investigate the role of ERCC1 in CSR, the nucleotide sequences of switch (S) regions were determined. The mutation frequency in germline Sμ segments and recombined Sμ-Sγ3 segments cloned from Ercc1−/− splenic B cells induced to switch in culture was identical to that of wild-type (WT) littermates. However, Ercc1−/− cells show increased targeting of the mutations to G:C bp in RGYW/WRCY hotspots and mutations occur at sites more distant from the S–S junctions compared with WT mice. The results indicate that ERCC1 is not epistatic with MMR and suggest that ERCC1 might be involved in processing or repair of DNA lesions in S regions during CSR

A link between the accumulation of DNA damage and loss of multi-potency of human mesenchymal stromal cells

Human mesenchymal stromal cells (hMSCs) represent an attractive cell source for clinic applications. Besides being multi-potent, recent clinical trials suggest that they secrete both trophic and immunomodulatory factors, allowing allogenic MSCs to be used in a wider variety of clinical situations. The yield of prospective isolation is however very low, making expansion a required step toward clinical applications. Unfortunately, this leads to a significant decrease in their stemness. To identify the mechanism behind loss of multi-potency, hMSCs were expanded until replicative senescence and the concomitant molecular changes were characterized at regular intervals. We observed that, with time of culture, loss of multi-potency was associated with both the accumulation of DNA damage and the respective activation of the DNA damage response pathway, suggesting a correlation between both phenomena. Indeed, exposing hMSCs to DNA damage agents led to a significant decrease in the differentiation potential. We also showed that hMSCs are susceptible to accumulate DNA damage upon in vitro expansion, and that although hMSCs maintained an effective nucleotide excision repair activity, there was a progressive accumulation of DNA damage. We propose a model in which DNA damage accumulation contributes to the loss of differentiation potential of hMSCs, which might not only compromise their potential for clinical applications but also contribute to the characteristics of tissue agein

LOUPE: Observing Earth from the Moon to prepare for detecting life on Earth-like exoplanets

LOUPE, the Lunar Observatory for Unresolved Polarimetry of the Earth, is a small, robust spectro-polarimeter with a mission to observe the Earth as an exoplanet. Detecting Earth-like planets in stellar habitable zones is one of the key challenges of modern exoplanetary science. Characterising such planets and searching for traces of life requires the direct detection of their signals. LOUPE provides unique spectral flux and polarisation data of sunlight reflected by the Earth, the only planet known to harbor life. This data will be used to test numerical codes to predict signals of Earth-like exoplanets, to test algorithms that retrieve planet properties, and to fine-tune the design and observational strategies of future space observatories. From the Moon, LOUPE will continuously see the entire Earth, enabling it to monitor the signal changes due to the planet's daily rotation, weather patterns, and seasons, across all phase angles. Here, we present both the science case and the technology behind LOUPE's instrumental and mission design.Comment: 13 pages, 5 figures. Accepted for publication in Royal Society Philosophical Transactions A. Corrected typos in v

DNA damage stabilizes interaction of CSB with the transcription elongation machinery

The Cockayne syndrome B (CSB) protein is essential for transcription-coupled DNA repair (TCR), which is dependent on RNA polymerase II elongation. TCR is required to quickly remove the cytotoxic transcription-blocking DNA lesions. Functional GFP-tagged CSB, expressed at physiological levels, was homogeneously dispersed throughout the nucleoplasm in addition to bright nuclear foci and nucleolar accumulation. Photobleaching studies showed that GFP-CSB, as part of a high molecular weight complex, transiently interacts with the transcription machinery. Upon (DNA damage-induced) transcription arrest CSB binding these interactions are prolonged, most likely reflecting actual engagement of CSB in TCR. These findings are consistent with a model in which CSB monitors progression of transcription by regularly probing elongation complexes and becomes more tightly associated to these complexes when TCR is active