Hi! You reached the research group of Jernej Ule, based at the Department of Molecular Neuroscience at the UCL Institute of Neurology.

  • UCL lab website
  • UCL research website

  • Why RNA?

    The primary task of the nervous system is to process and store information, and we study how this is achieved at the level of RNA molecules. Each RNA passes through several regulatory stages, which are controlled by protein-RNA complexes. In addition to the regulated pre-mRNA or mRNA, these complexes include non-coding RNAs (ncRNAs), RNA-binding proteins (RBPs) and other associated proteins, which together form ribonucleoprotein complexes (RNPs).

    We study the structure and function of protein-RNA complexes in the brain. In recent years, we have studied how these complexes regulate alternative splicing, and how aberrant function of protein-RNA complexes contribute to neurologic diseases. Our ultimate goal is to understand how the sequence and structure of RNAs defines the dynamic composition and function of RNPs. We develop innovative transcriptomic techniques such as UV cross-linking and immunoprecipitation (iCLIP) to study protein-RNA interactions in intact cells on a transcriptomic level. We also develop computational approaches to identify new regulatory mechanisms by integrating data produced by multiple high-throughput methods, and then explore these mechanisms in detail using cellular and molecular biology.

    We culture induced pluripotent stem cells from healthy individuals or patients with disease-causing mutations to examine how specific genetic mutations cause motor neuron disease. To understand the mechanisms behind these mutations, we modify the cells further with genome editing and differentiate them into specific neuronal or glial cell types. This allows us to study how the composition of RNPs changes in response to cellular signals, such as neuronal synaptic activity or the signals that initiate motor neuron disease. Specifically, we aim to:

    1) Determine how the structure of protein-RNA complexes instructs their function in neurons.

    2) Understand how protein-RNA complexes respond to neuronal signals, particularly in motor neurons.

    3) Define how mutations that affect the composition of protein-RNA complexes can impact RNA regulation in neurons, and thereby contribute to neurologic diseases.

    An RNA molecule passes through many compartments in the cell, where it undergoes processing, editing or sophisticated regulation. And here are some of the RNA stories that we have passed through:


    Developing the techniques to study protein-RNA interactions.

    Methods for nucleotide-resolution studies of protein-RNA interactions We developed individual-nucleotide resolution UV crosslinking and immunoprecipitation (iCLIP) to quantify protein-RNA interactions in the whole transcriptome. We review the progress made in the last years in the technologies for studies of protein-RNA interactions. You can download the manuscript here. We also performed a comparative analysis of iCLIP and CLIP, click here.

    RNA maps show how the function of RBP is defined by the location of its RNA binding site.

    Image of the decision-making process of the splicesome as it scans the pre-mRNA for exons, and the role of RNA-binding proteins in modulating this process. We integrate transcriptomic data on protein-RNA interactions and their function, which can tell us how ribonucleoproteins (RNPs) assemble at specific positions on their target transcripts and thereby regulate alternative splicing, mRNA decay or translation. For example, we used iCLIP to assess where an RBP binds its target transcripts, and RNA-seq to assess how this RBP controls pre-mRNA processing. This approach revealed that most RBPs regulate alternative splicing according to genome-wide positional principles, or RNA splicing maps. For example. by integrating TIA iCLIP with splicing analysis upon TIA knockdown, we were able to derive nucleotide-resolution RNA splicing maps of TIA proteins. Moreover, we developed software (RNAmotifs) that can derive RNA splicing maps by analysis of multivalent RNA motifs that are often bound by RBPs.

    Understanding the regulation and function of cryptic splicing elements.

    Alu-derived exons

    Artistic rendering of ultraviolet crosslinking of the hnRNP C1/C2 tetramer interacting with pre-mRNA. By identifying RNA binding sites of hnRNP C across the transcriptome, we have shown that hnRNP C specifically recognizes long uridine tracts, and can thereby repress splicing of alternative exons. This was evident by the ultraviolet crosslinking of the hnRNP C1/C2 tetramer, which demonstrated that hnRNP C forms higher-order complexes that bind across the repressed exons (see the paper). This uncovered a major role for hnRNP C in the repression of cryptic splicing elements(see the paper). We found that hnRNP C controls the emergence of new exons from Alu elements, which are retrotransposable elements that are specific for primate genomes, and constitute 10% of human genome. hnRNP C represses recognition of cryptic splice sites in Alu elements by displacing the splicing factor U2AF65 from uridine tracts. Loss of hnRNP C leads to formation of thousands of harmful exons, and mutations disrupting hnRNP C binding cause human diseases. Since the repressive function of hnRNP C prevents the damaging effects of immediate Alu exonization, it enables mutations to gradually create Alu-derived exons. This represents an elegant molecular mechanism that could mediate incremental evolution of new cellular functions.

    Recursive splicing in long introns

    Gene - recursive splicing - mRNA - brain, by Petra Kokol Long introns contain hundreds of so-called ‘cryptic sequences’ that appear very similar to exons, but are not supposed to be used. The cellular machinery faces great challenges in distinguishing true exons from these cryptic sites. We found that cells sometimes select a cryptic exon that is present deep within a long intron, but later discard it, in a process called recursive splicing (see the paper here). Normally recursive exon removes this cryptic exon, allowing it to remain invisible. However, if the recursive site is preceded by other cryptic splicing events, then the exon is not removed – creating a ‘binary switch’ or checkpoint that can distinguish correct splicing events from the newly emerging cryptic events, which could be potentially damaging. Thus, long introns on one hand enable emergence of many cryptic splicing events during evolution, whereas recursive splicing ensures that this evolutionary tinkering does not disturb the primary mRNA that needs to be made from the gene. We observed this process happening in some of the longest genes that are expressed in human brain, which are often implicated in autism or other neurodevelopmental disorders.

    Understanding the secondary structure of full-length mRNAs, and its role in RNP assembly.

    Lines between the two arms of RNA duplexes The secondary structure of mRNAs has important effects on its stability and translation. To understand the in vivo structure of full-length mRNAs, developed a technique called hiCLIP to identify the connections that hook sections of an mRNA together, which are called RNA duplexes. We were amazed to find that mRNAs form thousands of such duplexes, and often these duplexes hook together very distant parts of mRNA molecules. We found that that these duplexes interact with the double-stranded RNA binding protein Staufen 1. We also found that these RNA duplexes have less genetic variation in humans than surrounding areas of the mRNA, indicating that mutations could cause disease by disrupting the structure of mRNAs. See the paper here.

    Understanding the role of RNPs in brain function and disease.

    Alternative splicing can produce several mRNA isoforms from a gene, and these isoforms can change in the human brain during aging or neurodegeneration (click here). Moreover, we uncovered the regulatory networks controlled by TDP-43 and FUS, two proteins can cause amyotrophic lateral sclerosis when mutated. We showed that both proteins regulate a functionally coherent set of transcripts, many of which encode proteins implicated in neurodegenerative disorders click here or here).