The mechanisms of a mammalian splicing enhancer

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Jobbins, Andrew M and Reichenbach, Linus F and Lucas, Christian M and Hudson, Andrew J and Burley, Glenn A and Eperon, Ian C (2018) The mechanisms of a mammalian splicing enhancer. Nucleic Acids Research. ISSN 1362-4962 (https://doi.org/10.1093/nar/gky056)

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Abstract

Exonic splicing enhancer (ESE) sequences are bound by serine & arginine-rich (SR) proteins, which in turn enhance the recruitment of splicing factors. It was inferred from measurements of splicing around twenty years ago that Drosophila doublesex ESEs are bound stably by SR proteins, and that the bound proteins interact directly but with low probability with their targets. However, it has not been possible with conventional methods to demonstrate whether mammalian ESEs behave likewise. Using single molecule multi-colour colocalization methods to study SRSF1-dependent ESEs, we have found that that the proportion of RNA molecules bound by SRSF1 increases with the number of ESE repeats, but only a single molecule of SRSF1 is bound. We conclude that initial interactions between SRSF1 and an ESE are weak and transient, and that these limit the activity of a mammalian ESE. We tested whether the activation step involves the propagation of proteins along the RNA or direct interactions with 3′ splice site components by inserting hexaethylene glycol or abasic RNA between the ESE and the target 3′ splice site. These insertions did not block activation, and we conclude that the activation step involves direct interactions. These results support a model in which regulatory proteins bind transiently and in dynamic competition, with the result that each ESE in an exon contributes independently to the probability that an activator protein is bound and in close proximity to a splice site.

ORCID iDs

Jobbins, Andrew M, Reichenbach, Linus F ORCID logoORCID: https://orcid.org/0000-0002-9977-0659, Lucas, Christian M, Hudson, Andrew J, Burley, Glenn A and Eperon, Ian C;
  • Item type:Article
    ID code:63183
    Dates:
    Date
    Event
    31 January 2018
    Published
    19 January 2018
    Accepted
    Notes:This paper was classed as a Breakthrough Article. NAR’s Breakthrough Articles present high-impact studies answering long-standing questions in the field of nucleic acids research and/or opening up new areas and mechanistic hypotheses for investigation. These articles are chosen by the Editors on the recommendation of Editorial Board Members and Referees. Articles are accompanied by a brief synopsis explaining the findings of the paper and where they fit in the broader context of nucleic acids research. They represent the very best papers published at NAR.
    Subjects:Science > Chemistry
    Department:Faculty of Science > Pure and Applied Chemistry
    Technology and Innovation Centre > Bionanotechnology
    Depositing user: Pure Administrator
    Date deposited:05 Feb 2018 12:43
    Last modified:11 Nov 2024 11:55
    Related URLs:
    URI:https://strathprints.strath.ac.uk/id/eprint/63183
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