Structural studies of the human ribosome involved in ribosomopathies

Ribosomes are the cellular machines for translating genetic code to proteins in all living cells, hence essential for cell survival, cellular growth and division. Here we focus on a ribosomopathy genetic syndrome named Diamond Blackfan Anemia (DBA), which is connected to single mutations in any of 20 rProtein genes. Based on initial bioinformatic studies, we gained preliminary suggestions on consequent effects of a few of these mutations on ribosome`s structure as well as on cellular and molecular processes. However, owing to the lack of structural data, these mutations` structural and molecular implications are still not confirmed. Assuming that some of these mutations are indeed expressed and the mutated proteins are incorporated in the patient’s ribosomes, we expect to shed light on the molecular basis for these diseases by detrmining the structures of these ribosomes using cryo-EM techniques. For this aim we purified mutated ribosomes from a few cell types: I. Cells that were mutated by CRISPR-Cas9-based genome editing techniques that were applied in vitro on human Embryonic Stem cells (hESc). II. Cells originated from patients` bone marrow samples reprogrammed into induced Pluripotent Stem cells (iPSc). We expect that by determining the structures of these mutated ribosomes (and of the unmutated control ribosomes), we will shed light on the intriguing question of how a single mutation in a single protein of a huge molecular machine (m.w > 4.5 MDalton), the ribosome, can lead to a significant clinical effect. We aim to use these insights for therapeutic developments.