@phdthesis{Schuster2020, author = {Schuster, Maja}, title = {High resolution decoding of the tobacco chloroplast translatome and its dynamics during light-intensity acclimation}, doi = {10.25932/publishup-51268}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-512680}, school = {Universit{\"a}t Potsdam}, pages = {xvii, 155}, year = {2020}, abstract = {Chloroplasts are the photosynthetic organelles in plant and algae cells that enable photoautotrophic growth. Due to their prokaryotic origin, modern-day chloroplast genomes harbor 100 to 200 genes. These genes encode for core components of the photosynthetic complexes and the chloroplast gene expression machinery, making most of them essential for the viability of the organism. The regulation of those genes is predominated by translational adjustments. The powerful technique of ribosome profiling was successfully used to generate highly resolved pictures of the translational landscape of Arabidopsis thaliana cytosol, identifying translation of upstream open reading frames and long non-coding transcripts. In addition, differences in plastidial translation and ribosomal pausing sites were addressed with this method. However, a highly resolved picture of the chloroplast translatome is missing. Here, with the use of chloroplast isolation and targeted ribosome affinity purification, I generated highly enriched ribosome profiling datasets of the chloroplasts translatome for Nicotiana tabacum in the dark and light. Chloroplast isolation was found unsuitable for the unbiased analysis of translation in the chloroplast but adequate to identify potential co-translational import. Affinity purification was performed for the small and large ribosomal subunit independently. The enriched datasets mirrored the results obtained from whole-cell ribosome profiling. Enhanced translational activity was detected for psbA in the light. An alternative translation initiation mechanism was not identified by selective enrichment of small ribosomal subunit footprints. In sum, this is the first study that used enrichment strategies to obtain high-depth ribosome profiling datasets of chloroplasts to study ribosome subunit distribution and chloroplast associated translation. Ever-changing light intensities are challenging the photosynthetic capacity of photosynthetic organism. Increased light intensities may lead to over-excitation of photosynthetic reaction centers resulting in damage of the photosystem core subunits. Additional to an expensive repair mechanism for the photosystem II core protein D1, photosynthetic organisms developed various features to reduce or prevent photodamage. In the long-term, photosynthetic complex contents are adjusted for the efficient use of experienced irradiation. However, the contribution of chloroplastic gene expression in the acclimation process remained largely unknown. Here, comparative transcriptome and ribosome profiling was performed for the early time points of high-light acclimation in Nicotiana tabacum chloroplasts in a genome-wide scale. The time- course data revealed stable transcript level and only minor changes in translational activity of specific chloroplast genes during high-light acclimation. Yet, psbA translation was increased by two-fold in the high light from shortly after the shift until the end of the experiment. A stress-inducing shift from low- to high light exhibited increased translation only of psbA. This study indicate that acclimation fails to start in the observed time frame and only short-term responses to reduce photoinhibition were observed.}, language = {en} } @misc{BenteleSaffertRauscheretal.2013, author = {Bentele, Kajetan and Saffert, Paul and Rauscher, Robert and Ignatova, Zoya and Bluethgen, Nils}, title = {Efficient translation initiation dictates codon usage at gene start}, series = {Postprints der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, journal = {Postprints der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, number = {912}, issn = {1866-8372}, doi = {10.25932/publishup-44133}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-441337}, pages = {12}, year = {2013}, abstract = {The genetic code is degenerate; thus, protein evolution does not uniquely determine the coding sequence. One of the puzzles in evolutionary genetics is therefore to uncover evolutionary driving forces that result in specific codon choice. In many bacteria, the first 5-10 codons of protein-coding genes are often codons that are less frequently used in the rest of the genome, an effect that has been argued to arise from selection for slowed early elongation to reduce ribosome traffic jams. However, genome analysis across many species has demonstrated that the region shows reduced mRNA folding consistent with pressure for efficient translation initiation. This raises the possibility that unusual codon usage is a side effect of selection for reduced mRNA structure. Here we discriminate between these two competing hypotheses, and show that in bacteria selection favours codons that reduce mRNA folding around the translation start, regardless of whether these codons are frequent or rare. Experiments confirm that primarily mRNA structure, and not codon usage, at the beginning of genes determines the translation rate.}, language = {en} } @misc{EcksteinSchwarz2018, author = {Eckstein, Lars and Schwarz, Anja}, title = {The Making of Tupaia's Map}, series = {Postprints der Universit{\"a}t Potsdam Philosophische Reihe}, journal = {Postprints der Universit{\"a}t Potsdam Philosophische Reihe}, number = {154}, issn = {1866-8380}, doi = {10.25932/publishup-42309}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-423091}, pages = {96}, year = {2018}, abstract = {Tupaia's Map is one of the most famous and enigmatic artefacts to emerge from the early encounters between Europeans and Pacific Islanders. It was drawn by Tupaia, an arioi priest, chiefly advisor and master navigator from Ra'iātea in the Leeward Society Islands in collaboration with various members of the crew of James Cook's Endeavour, in two distinct moments of mapmaking and three draft stages between August 1769 and February 1770. To this day, the identity of many islands on the chart, and the logic of their arrangement have posed a riddle to researchers. Drawing in part on archival material hitherto overlooked, in this long essay we propose a new understanding of the chart's cartographic logic, offer a detailed reconstruction of its genesis, and thus for the first time present a comprehensive reading of Tupaia's Map. The chart not only underscores the extent and mastery of Polynesian navigation, it is also a remarkable feat of translation between two very different wayfinding systems and their respective representational models.}, language = {en} } @misc{LukoszekFeistIgnatova2016, author = {Lukoszek, Radoslaw and Feist, Peter and Ignatova, Zoya}, title = {Insights into the adaptive response of Arabidopsis thaliana to prolonged thermal stress by ribosomal profiling and RNA-Seq}, series = {BMC plant biology}, journal = {BMC plant biology}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-407262}, pages = {13}, year = {2016}, abstract = {Background: Environmental stress puts organisms at risk and requires specific stress-tailored responses to maximize survival. Long-term exposure to stress necessitates a global reprogramming of the cellular activities at different levels of gene expression. Results: Here, we use ribosome profiling and RNA sequencing to globally profile the adaptive response of Arabidopsis thaliana to prolonged heat stress. To adapt to long heat exposure, the expression of many genes is modulated in a coordinated manner at a transcriptional and translational level. However, a significant group of genes opposes this trend and shows mainly translational regulation. Different secondary structure elements are likely candidates to play a role in regulating translation of those genes. Conclusions: Our data also uncover on how the subunit stoichiometry of multimeric protein complexes in plastids is maintained upon heat exposure.}, language = {en} }