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Investigating the Effects of Autocleavage of Yeast Spliceosomal Protein Dib1
Author(s) -
Jamison Danielle,
Bullock Zoe,
Meeks Garrison,
Buergler Eva,
Maeder Corina
Publication year - 2021
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2021.35.s1.03645
Subject(s) - spliceosome , rna splicing , snrnp , chemistry , intron , yeast , rna , biochemistry , computational biology , gene , biology
Splicing, the removal of non‐coding introns and ligation of coding exons from pre‐messenger RNA (pre‐mRNA), is an essential component of eukaryotic life. The spliceosome facilitates splicing and is composed of five small nuclear ribonucleoproteins (snRNPs) and over 100 associated proteins. Dib1 is a critical protein involved in the transition of the spliceosome from pre‐catalytic to catalytic so the spliceosome can perform the two required trans‐esterification steps. The human ortholog Dim1 demonstrates autocleavage, or the removal of the last 14 amino acids from its C‐terminal tail. Preliminary results from my research suggest that yeast ortholog Dib1 also demonstrates this activity, autocleaving 13 instead of 14 amino acids from its C‐terminal tail. Determining the biological significance of this mechanism may provide insight into a new regulatory mechanism of the spliceosome. To study the autocleavage reaction, we have purified yeast Dib1 and human Dim1 proteins and performed a series of auto‐cleavage reactions. Progress is monitored periodically over a time span of two weeks and mass spectrometry is used to confirm autocleavage. Results showing the importance of divalent cations, particularly zinc, in inducing autocleavage in Dib1 will be presented. Limitations and future directions for the project will also be presented.