Characterizing the Role of p14 in Pre-mRNA Splicing

  • Author / Creator
    Madi, Sayed
  • Splicing is an essential step in the post-transcriptional modification of pre-mRNAs in eukaryotes. This process is catalyzed by the spliceosome, a multi-megadalton nuclear protein-RNA enzyme which excises non-coding sequence from RNA transcripts prior to translation. Deficient or improper splicing of introns in humans has been linked to many morbidities including myelodysplastic syndromes (MDS) and retinitis pigmentosa.

    SF3b14a/p14 is an evolutionarily conserved 14 kDa protein within the spliceosome that has been shown to crosslink to a highly conserved nucleotide at the 3′ end of introns that performs the first step of splicing. To date, the exact function that p14 plays in splicing remains elusive. Interestingly, siRNA knockdown of p14 in HeLa cells results in cell death within a few days, therefore making it an essential gene in humans.

    I have used the model organism Schizosaccharomyces pombe to study the role of p14 in pre-mRNA splicing. This was possible because the S. pombe p14 knockout is viable. Therefore, by investigating differences between wild-type and p14 knockout cells, I hoped to gather new insights into protein function. To do this, I utilized a combination of stress tests, morphological studies, and mass spectrometry.

    My research has shown that p14 may play an important role in the recognition and subsequent splicing of a certain subset of introns. This was supported by mass spectrometry data showing a downregulation in proteins required for organo-nitrogen compound metabolism and mitosis in p14 knockout cells.

  • Subjects / Keywords
  • Graduation date
    Spring 2021
  • Type of Item
  • Degree
    Master of Science
  • DOI
  • License
    This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.