Cryo-EM reconstructions of translation complexes
Abstract
We are structural biologists interested in translation, the cellular process where mRNA sequence is translated into amino acid polymers, i.e. proteins. This process is localized to the ribosomes, which are huge macromolecular complexes that consists of RNA and proteins. In this application, we are requesting GPU resources for processing of electron microscopy data for structure determination of translation-related complexes.
In all kingdoms of life, the ribosomal RNA is posttranscriptionally modified, which provide evolutionary advantages under specific conditions. In bacteria, the rRNA modifications are added by site-specific enzymes. In on-going projects, we aim to elucidate how rRNA modification enzymes involved in ribosome biogenesis and antibiotic resistance specifically recognize their target nucleotides, how they catalyze the reaction and how the modification influences the structure, stability and function of the local RNA region as well as the mature ribosome. In another project, we study the molecular mechanisms of bacterial resistance to the antibiotic fusidic acid (FA). FA targets both the translocation and the ribosome-recycling stage of translation. FusB-type resistance is mediated by the resistance protein FusB, that binds with high affinity to the FA target, elongation factor G. We aim to use cryo-EM to further clarify this mechanism of resistance. In another project, we aim to clarify ribosome recycling and how FA inhibits this process.
We are collecting cryo-EM data at the SciLifeLab National Facilities for Cryo-EM in Solna and Umeå and at the Diamond eBIC facility. In addition, our department runs a cryo-EM facility at Uppsala University, where we also will collect data.
