A5014386933- RNA modifications and cancer
- RNA and protein synthesis mechanisms
- RNA Research and Splicing
- Peptidase Inhibition and Analysis
- Ubiquitin and proteasome pathways
- Cancer-related Molecular Pathways
- Virus-based gene therapy research
- Cancer-related gene regulation
- CRISPR and Genetic Engineering
- Viral gastroenteritis research and epidemiology
- DNA Repair Mechanisms
- Advanced biosensing and bioanalysis techniques
- T-cell and Retrovirus Studies
- Bacteriophages and microbial interactions
- Molecular Biology Techniques and Applications
- Epigenetics and DNA Methylation
- Cytomegalovirus and herpesvirus research
- Inorganic Fluorides and Related Compounds
CSL (Switzerland)
2020-2024
ETH Zurich
2012-2022
Life Science Zurich
2014-2015
Ribosome biogenesis is a highly complex process requiring many assisting factors. Studies in yeast have yielded comprehensive knowledge of the cellular machinery involved this process. However, aspects ribosome synthesis are different higher eukaryotes, and global set mammalian factors remains unexplored. We used an imaging-based, genome-wide RNAi screen to find human proteins 40S ribosomal subunit biogenesis. Our analysis identified ∼300 factors, part essential protein modules such as small...
The nucleolar surveillance pathway monitors integrity and responds to stress by mediating binding of ribosomal proteins MDM2, resulting in p53 accumulation. Inappropriate activation is implicated the pathogenesis ribosomopathies, while drugs selectively activating are trials for cancer. Despite this, molecular mechanism(s) regulating this process poorly understood. Using genome-wide loss-of-function screens, we demonstrate ribosome biogenesis axis as most potent class genes whose disruption...
Abstract The DNA-PKcs inhibitor AZD7648 enhances CRISPR–Cas9-directed homology-directed repair efficiencies, with potential for clinical utility, but its possible on-target consequences are unknown. We found that genome editing causes frequent kilobase-scale and megabase-scale deletions, chromosome arm loss translocations. These large-scale chromosomal alterations evade detection through typical assays, prompting caution in deploying reinforcing the need to investigate multiple types of outcomes.
Synthesis of ribosomal subunits in eukaryotes is a complex and tightly regulated process that has been mostly characterized yeast. The discovery growing number diseases linked to defects ribosome biogenesis calls for deeper understanding these mechanisms the specificities human maturation. We present 19 Å resolution cryo-EM reconstruction cytoplasmic precursor small subunit, purified by using tagged factor LTV1 as bait. Compared yeast pre-40S particles, this first three-dimensional structure...
The poly-A specific ribonuclease (PARN), initially characterized for its role in mRNA catabolism, supports the processing of different types non-coding RNAs including telomerase RNA. Mutations PARN are linked to dyskeratosis congenita and pulmonary fibrosis. Here, we show that is part enzymatic machinery matures human 18S ribosomal RNA (rRNA). Consistent with nucleolar steady-state localization, required 40S subunit production co-purifies precursors. Depletion or expression a...
Establishment of translational competence represents a decisive cytoplasmic step in the biogenesis 40S ribosomal subunits. This involves final 18S rRNA processing and release residual factors, including protein kinase RIOK1. To identify novel proteins promoting maturation human subunits, we characterized pre-ribosomal subunits trapped on RIOK1 by mass spectrometry, identified deubiquitinase USP16 among captured factors. We demonstrate that constitutes component late pre-40S promotes removal...
Preventing premature interaction of pre-ribosomes with the translation apparatus is essential for translational accuracy. Hence, final maturation step releasing functional 40S ribosomal subunits, namely processing 18S RNA 3′ end, safeguarded by protein DIM2, which both interacts endoribonuclease NOB1 and masks rRNA cleavage site. To elucidate control mechanism that unlocks activity, we performed cryo-electron microscopy analysis late human pre-40S particles purified using a catalytically...
Biogenesis of 40S pre-ribosomal subunits requires many trans-acting factors, among them several protein kinases. In this study we show that the human CK1 isoforms δ and ε are required for cytoplasmic maturation steps subunit precursors. We both components pre-40S subunits, where they phosphorylate ribosome biogenesis factors ENP1/BYSL LTV1. inhibition or co-depletion CK1δ result in failure to recycle a series including ENP1/BYSL, LTV1, RRP12, DIM2/PNO1, RIO2 NOB1 from particles after nuclear...
The interleukin enhancer binding factors ILF2 (NF45) and ILF3 (NF90/NF110) have been implicated in various cellular pathways, such as transcription, microRNA (miRNA) processing, DNA repair, translation, mammalian cells. Using tandem affinity purification, we identified human NF45 NF90 components of precursors to 60S (pre-60S) ribosomal subunits. are enriched nucleoli cosediment with pre-60S particles density gradient analysis. We show that association the NF45/NF90 heterodimer requires...
BRCA2 and CDKN1A(p21,CIP1)‐interacting protein (BCCIP) is an evolutionary conserved implicated in maintenance of genome stability cell cycle progression. Two isoforms BCCIP with distinct C‐terminal domains exist humans. We show that mammalian BCCIPβ, but not BCCIPα, forms a ternary complex the ribosomal RPL23/uL14 pre‐60S trans‐acting factor eIF6. Complex formation dependent on intact domain BCCIPβ. Depletion BCCIPβ reduces pool free RPL23, decreases eIF6 levels nucleoli. Overexpression...
Despite recent advances made toward improving the efficacy of lentiviral gene therapies, a sizeable proportion produced vector contains an incomplete and thus potentially nonfunctional RNA genome. This can undermine delivery by lentivirus as well increase manufacturing costs must be improved to facilitate widespread clinical implementation therapies. Here, we compare three long-read sequencing technologies for their ability detect issues in design determine nanopore direct most powerful. We...
Despite recent advances made towards improving the efficacy of lentiviral gene therapies, a sizeable proportion produced vector contains an incomplete and thus potentially non-functional RNA genome. This can undermine delivery by lentivirus as well increase manufacturing costs must be improved to facilitate widespread clinical implementation therapies. Here, we compare three long-read sequencing technologies for their ability detect issues in design determine Nanopore direct most powerful....
Abstract The nucleolar surveillance pathway (NSP) monitors fidelity and responds to stresses (i.e., inactivation of ribosome biogenesis) by mediating the inhibitory binding ribosomal proteins (RPs) mouse double minute 2 homolog (MDM2), a nuclear-localised E3 ubiquitin ligase, which results in p53 accumulation. Inappropriate activation NSP has been implicated pathogenesis collection human diseases termed “ribosomopathies”, while drugs that selectively activate are now trials for cancer....
ABSTRACT Preventing premature interaction of preribosomes with the translation apparatus is essential to accuracy. Hence, final maturation step releasing functional 40S ribosomal subunits, namely processing 18S RNA 3’ end, safeguarded by protein DIM2, which both interacts endoribonuclease NOB1 and masks rRNA cleavage site. To elucidate control mechanism that unlocks activity, we performed cryo-EM analysis late human pre-40S particles purified using a catalytically-inactive form ATPase RIO1....