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Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes
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Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes

Published on: November 9, 2020

Nuclear pre-mRNA splicing in the fission yeast Schizosaccharomyces pombe strictly requires an intron-contained,

P Mertins, D Gallwitz

    The EMBO Journal
    |June 1, 1987
    PubMed

    Insights

    The fission yeast Schizosaccharomyces pombe splicing mechanism is not similar to metazoans. A conserved sequence element, 5'-CTPuAPy-3', is essential for efficient splicing and branch formation in S. pombe.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Yeast Biology

    Background:

    • Recent arguments suggested pre-mRNA splicing in Schizosaccharomyces pombe resembles metazoan splicing more than Saccharomyces cerevisiae splicing.
    • This study investigates the conserved sequence elements and mechanisms involved in S. pombe intron splicing.

    Purpose of the Study:

    • To challenge the assumption of similarity between S. pombe and metazoan splicing.
    • To identify and characterize the essential conserved sequence elements in S. pombe introns.
    • To elucidate the role of these elements in the splicing and branch formation process.

    Main Methods:

    • Sequence analysis of S. pombe introns to identify conserved elements.
    • Site-directed mutagenesis of the conserved 5 ahydro-CTPuAPy-3 ahydro sequence.
    • Functional analysis of splicing efficiency in S. pombe.
    • Complementation experiments using a Saccharomyces cerevisiae intron in S. pombe.

    Main Results:

    • The conserved sequence element 5 ahydro-CTPuAPy-3 ahydro, located upstream of the 3 ahydro splice site in S. pombe introns, is indispensable for efficient splicing.
    • The adenine residue within the 5 ahydro-CTPuAPy-3 ahydro sequence is crucial for branch formation.
    • Point mutations in the 5 ahydro-CTPuAPy-3 ahydro sequence abolish splicing and do not lead to cryptic branch site usage.
    • A Saccharomyces cerevisiae intron is efficiently spliced in S. pombe, utilizing its TACTAAC box for branch formation.

    Conclusions:

    • Contrary to recent arguments, S. pombe pre-mRNA splicing is not more similar to metazoan splicing than to S. cerevisiae splicing.
    • The conserved 5 ahydro-CTPuAPy-3 ahydro sequence element plays a critical role in S. pombe splicing, analogous to the TACTAAC box in S. cerevisiae.
    • S. pombe possesses the machinery to correctly splice introns from S. cerevisiae, highlighting conserved splicing mechanisms.

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