A5001482627- RNA and protein synthesis mechanisms
- RNA modifications and cancer
- RNA Research and Splicing
- Mitochondrial Function and Pathology
- Nuclear Structure and Function
- Biochemical Acid Research Studies
- Genomics and Chromatin Dynamics
- Signaling Pathways in Disease
- Ubiquitin and proteasome pathways
- Peptidase Inhibition and Analysis
- DNA Repair Mechanisms
- Enzyme Structure and Function
- ATP Synthase and ATPases Research
- Microbial Metabolic Engineering and Bioproduction
- Trypanosoma species research and implications
- Mycobacterium research and diagnosis
- Molecular Biology Techniques and Applications
- Bacterial Genetics and Biotechnology
- Advanced Electron Microscopy Techniques and Applications
- Amino Acid Enzymes and Metabolism
- Microtubule and mitosis dynamics
- Inorganic Fluorides and Related Compounds
- Metabolism and Genetic Disorders
- Electron and X-Ray Spectroscopy Techniques
- DNA and Nucleic Acid Chemistry
Science for Life Laboratory
2016-2024
Stockholm University
2017-2024
AstraZeneca (United Kingdom)
2023
Max Planck Institute for Biophysical Chemistry
2020-2021
MRC Laboratory of Molecular Biology
2015-2017
Medical Research Council
2015-2016
To initiate cotranscriptional splicing, RNA polymerase II (Pol II) recruits the U1 small nuclear ribonucleoprotein particle (U1 snRNP) to nascent precursor messenger (pre-mRNA). Here, we report cryo-electron microscopy structure of a mammalian transcribing Pol II-U1 snRNP complex. The reveals that and interact directly. This interaction positions pre-mRNA 5' splice site near exit II. Extension retains site, leading formation "growing intron loop." Loop may facilitate scanning for 3'...
Translation of mitochondrial messenger RNA (mt-mRNA) is performed by distinct mitoribosomes comprising at least 36 mitochondria-specific proteins. How these mitoribosomal proteins assist in the binding mt-mRNA and to what extent they are involved translocation transfer (mt-tRNA) unclear. To visualize process translation human mitochondria, we report ~3.0 Å resolution structure mitoribosome, including L7/L12 stalk, eight structures its functional complexes with mt-mRNA, mt-tRNAs, recycling...
Abstract The mitoribosome translates mitochondrial mRNAs and regulates energy conversion that is a signature of aerobic life forms. We present 2.2 Å resolution structure human together with validated mitoribosomal RNA (rRNA) modifications, including aminoacylated CP-tRNA Val . shows how proteins stabilise binding mRNA tRNA helping to align it in the decoding center, whereas GDP-bound mS29 stabilizes intersubunit communication. Comparison between different states, respect position, allowed us...
Abstract Formation of 100S ribosome dimer is generally associated with translation suppression in bacteria. Trans -acting factors modulation factor (RMF) and hibernating promoting (HPF) were shown to directly mediate this process E. coli . Gram-positive S. aureus lacks an RMF homolog the structural basis for its formation was not known. Here we report cryo-electron microscopy structure native from , revealing molecular mechanism formation. The distinct previously reported analogs relies on...
Human ALC1 is an oncogene-encoded chromatin-remodeling enzyme required for DNA repair that possesses a poly(ADP-ribose) (PAR)-binding macro domain. Its engagement with PARylated PARP1 activates at sites of damage, but the underlying mechanism remains unclear. Here, we establish dual role domain in autoinhibition ATPase activity and coupling to nucleosome mobilization. In absence inactive conformation maintained by juxtaposition against predominantly C-terminal lobe through conserved...
Glioblastoma stem cells (GSCs) resist current glioblastoma (GBM) therapies. GSCs rely highly on oxidative phosphorylation (OXPHOS), whose function requires mitochondrial translation. Here we explore the therapeutic potential of targeting translation and report results high-content screening with putative blockers ribosomes. We identify bacterial antibiotic quinupristin/dalfopristin (Q/D) as an effective suppressor GSC growth. Q/D also decreases clonogenicity in vitro, consequently...
Mitoribosomes consist of ribosomal RNA and protein components, coordinated assembly which is critical for function. We used mitoribosomes from Trypanosoma brucei with reduced increased mass to provide insights into the biogenesis mitoribosomal large subunit. Structural characterization a stable intermediate revealed 22 factors, some have orthologues/counterparts/homologues in mammalian genomes. These factors form network that spans distance 180 Å, shielding surface. The central protuberance...
The NXF1:NXT1 complex (also known as TAP:p15) is a general mRNA nuclear export factor that conserved from yeast to humans. NXF1 modular protein constructed four domains (RRM, LRR, NTF2-like and UBA domains). It currently unclear how binds transcripts whether there higher organization of the domains. We report here 3.4 Å resolution crystal structure first three human together with NXT1 has two copies in asymmetric unit arranged form an intimate domain-swapped dimer. In this dimer, linkers...
Abstract There is a lack of current treatment options for ovarian clear cell carcinoma (CCC) and the cancer often resistant to platinum‐based chemotherapy. Hence there an urgent need novel therapeutics. The transcription factor hepatocyte nuclear 1β (HNF1β) ubiquitously overexpressed in CCC seen as attractive therapeutic target. This was validated through shRNA‐mediated knockdown target protein, HNF1β, five high‐ low‐HNF1β‐expressing lines. To inhibit protein function, cell‐permeable,...
RNA polymerase II (Pol II) facilitates co-transcriptional splicing by recruiting the U1 small nuclear ribonucleoprotein particle (U1 snRNP) to nascent transcripts. Here, we report cryo-electron microscopy structure of a transcribing Pol II-U1 snRNP complex with elongation factors DSIF and SPT6. Furthermore, our biochemical analysis revealed that phosphorylated carboxyl-terminal domain SPT6 interact directly proteins, facilitating its recruitment complex. This multivalent interaction allows...
Abstract Biogenesis of mitoribosomes requires dedicated chaperones, RNA-modifying enzymes, and GTPases, defects in mitoribosome assembly lead to severe mitochondriopathies humans. Here, we characterize late-step states the small mitoribosomal subunit (mtSSU) by combining genetic perturbation mutagenesis analysis with biochemical structural approaches. Isolation native mtSSU biogenesis intermediates via a FLAG-tagged variant GTPase MTG3 reveals three distinct states, which show how factors...
The mitoribosome translates mitochondrial mRNAs and regulates energy conversion that is a signature of aerobic life forms. We present 2.2 Å resolution structure human together with validated mitoribosomal RNA (rRNA) modifications, including aminoacylated CP-tRNA Val . shows how proteins stabilise binding mRNA tRNA helping to align it in the decoding center, whereas GDP-bound mS29 stabilizes intersubunit communication. Comparison between different states, respect position, allowed...
The TREX-2 complex integrates mRNA nuclear export into the gene expression pathway and is based on a Sac3 scaffold to which Thp1, Sem1, Sus1, Cdc31 bind. also binds factor, Mex67:Mtr2, through N-terminal region (Sac3N). Here, we characterize Chaetomium thermophilum TREX-2, show that in vitro reconstituted has an annular structure, define structural basis for interactions between Sac3, Cdc31, Mex67:Mtr2. Crystal structures binding of C. Sac3N Mex67 NTF2-like domain (Mex67NTF2L) mediated...
In Saccharomyces cerevisiae generation of export-competent mRNPs terminates the nuclear phase gene expression pathway and facilitates transport to cytoplasm for translation. Nab2 functions in this process control both mRNP compaction that movement through pore complexes length transcript poly(A) tails. has a modular structure includes seven CCCH Zn fingers bind A-rich RNAs 5–7 are critical these functions. Here, we demonstrate, using biophysical structural methods, binding A11G RNA induces...
Signaling through the receptor tyrosine kinase RET is essential during normal development. Both gain- and loss-of-function mutations are involved in a variety of diseases, yet molecular details activation have remained elusive. We reconstituted complete extracellular region signaling complex together with Neurturin (NRTN) GFRα2 determined its structure at 5.7-Å resolution by cryo-EM. The proteins form an assembly RET-GFRα2 RET-NRTN interfaces. Two key interaction points required for domain...
Abstract The pyruvate dehydrogenase complex (PDC) is a multienzyme central to aerobic respiration, connecting glycolysis mitochondrial oxidation of pyruvate. Similar the E3-binding protein (E3BP) mammalian PDC, PX selectively recruits E3 fungal but its divergent sequence suggests distinct structural mechanism. Here, we report reconstructions PDC from filamentous fungus Neurospora crassa by cryo-electron microscopy, where find X (PX) interior core as opposed substituting E2 subunits in...
Abstract The Mex67:Mtr2 complex is the principal yeast nuclear export factor for bulk mRNA and also contributes to ribosomal subunit export. Mex67 a modular protein constructed from four domains (RRM, LRR, NTF2-like UBA) that have been thought be joined by flexible linkers like beads on string, with RRM LRR binding RNAs UBA FG-nucleoporins facilitate movement through pores. Here, we show domain Saccharomyces cerevisiae RNA binding. Moreover, 3.3 Å resolution crystal structure of...
Significance The anaphase-promoting complex/cyclosome (APC/C) is a large E3 ubiquitin ligase that controls progression through mitosis and entry into G1. Its capacity to recognize ubiquitinate substrates dependent on coactivator subunits interact with substrate degrons promote conformational change of the APC/C increase its affinity for priming E2 UbcH10. We show WD40 domain complex subunit 1 (Apc1) required communicating initiated by binding catalytic module. In contrast UbcH10, elongating...
The human mitochondria possess a dedicated set of ribosomes (mitoribosomes) that translate 13 essential protein components the oxidative phosphorylation complexes encoded by mitochondrial genome. Since all proteins synthesized mitoribosomes are integral membrane proteins, tethered to inner during translation. Compared cytosolic ribosome mitoribosome has sedimentation coefficient 55S, half rRNA content, no 5S and 36 additional proteins. Therefore, higher protein-to-RNA ratio an atypical...
Abstract Eukaryotic gene transcription is carried out by three RNA polymerases: Pol I, II and III. Although it has long been known that I can form homodimers, unclear whether how the two other polymerases dimerize. Here we present cryo-electron microscopy (cryo-EM) structure of a mammalian dimer at 3.5 Å resolution. The differs from reveals one copy uses its RPB4-RPB7 stalk to penetrate active centre cleft copy, vice versa, giving rise molecular handshake. polymerase clamp domain displaced...