<> "The repository administrator has not yet configured an RDF license."^^ . <> . . "Proteomic Exploration of Protein-RNA Interactions in Human Cells"^^ . "The interactions of proteins with RNA are ubiquitous in human cell biology and fundamental\r\nto most if not all of life as we know it. Proteins and RNA interact for the purpose of gene\r\nexpression, where genetic information is encoded within RNA, which serves as template for\r\nprotein in messenger RNA, as scaffold for the trimming of other RNA within the spliceosome,\r\nas a catalyst for the production of protein within the ribosome, and as component of various\r\nother processes, many of which we are only beginning to understand. Central to our\r\nunderstanding of such processes is the interplay between proteins and RNA. Protein-RNA\r\ninteractions are often disrupted by conventional extraction methods and need to be stabilized\r\nthrough crosslinking in order to be appreciated by transcriptomic or proteomic methodology.\r\nCrosslinking typically occurs through UV irradiation, which covalently connects protein bound\r\nto RNA. However, currently there is no universal method for the extraction of proteincrosslinked\r\nRNA available, so that insights can only be gained for certain subsets of the\r\ntranscriptome or individual RNA-binding proteins, respectively. In this thesis a new method\r\nfor the extraction of protein-crosslinked RNA from UV-crosslinked cells is described. In order\r\nto demonstrate the comprehensive usefulness of the approach, which was termed XRNAX, it\r\nis applied for the resolution of several proteomic and transcriptomic problems. First, XRNAX\r\nis employed for the purification of ribonucleotide-crosslinked peptides identifying known and\r\nunknown protein-RNA interfaces. Next, a comprehensive draft for the human RNA-binding\r\nproteome is derived applying XRNAX to three commonly used cell lines. The same approach\r\nis applied for the differential quantification of RNA-binding during a timeline of arseniteinduced\r\ntranslational arrest in human cells. Additionally, XRNAX is combined with CLIPseq to\r\nmonitor the RNA exosome component EXOSC2 processing pre-ribosomal RNA during arsenite\r\nstress. At last, XRNAX is used along with TMT-SILAC in order to derive protein half-lives of the\r\nhuman RNA-bound proteome. Half-lives of the RNA-bound proteome are found on average\r\n1.75 fold increased in comparison to the total proteome. For validating the stabilization of\r\nprotein-RNA complexes, ribosomal assemblies are purified from human cells using polysome\r\nprofiling and protein half-lives assessed with TMT-SILAC. Stabilization is confirmed, however,\r\nonly for ribosomal proteins in 80S ribosomes within the 80S or polysome fractions. Further\r\nexperiments interfering with ribosome biogenesis, the proteasome and autophagy are\r\ndistilled into a model for the turnover of ribosomal proteins and ribosomal assemblies.\r\nIn summary, this thesis describes new methodology and biological insight on protein-RNA\r\ninteractions in human cells."^^ . "2019" . . . . . . . "Jakob"^^ . "Trendel"^^ . "Jakob Trendel"^^ . . . . . . "Proteomic Exploration of Protein-RNA Interactions in Human Cells (PDF)"^^ . . . "Trendel_Dissertation.pdf"^^ . . . "Proteomic Exploration of Protein-RNA Interactions in Human Cells (Other)"^^ . . . . . . "indexcodes.txt"^^ . . "HTML Summary of #26436 \n\nProteomic Exploration of Protein-RNA Interactions in Human Cells\n\n" . "text/html" . .