2026-09-19T13:45:59Zhttps://dspace.mit.edu/server/oai/requestoai:dspace.mit.edu:1721.1/1644892026-01-13T03:36:17Zcom_1721.1_7582com_1721.1_7581col_1721.1_131022
Burge, Christopher B.
Kenny, Connor Jens
Massachusetts Institute of Technology. Department of Biology
2026-01-12T19:40:03Z
2026-01-12T19:40:03Z
2025-09
2025-11-10T19:58:45.882Z
https://hdl.handle.net/1721.1/164489
Pre-mRNA splicing is an essential molecular process required for eukaryotic gene expression. In this thesis, I present a previously unknown mechanism of splicing regulation where a family of splicing factors, the LUC7 family, compete to differentially impact 5→ splice site (5→ SS) selection in a sequence-dependent manner. I quantitatively characterize two major subclasses of 5→ SS in eukaryotes and outline distinctive features of 5→ SS in exons affected by the three human LUC7 paralogs: LUC7L2 and LUC7L enhance splicing of “right-handed” 5→ SS that exhibit stronger consensus matching on the intron side of the nearly-invariant / GU, while LUC7L3 boosts splicing of “left-handed” 5→ SS with stronger consensus matching upstream of the /GU. Using a range of experimental systems, from human cells to mutant plants, I show that LUC7 paralogs have opposing effects on these two 5→ SS subclasses and that this regulatory mechanism likely originated in the last common ancestor of animals and plants over 1.5 billion years ago. I further evaluate a competing model of 5→ SS subclass regulation involving METTL16- mediated U6 snRNA modification and reconcile both models by devising computational tools that identify sequence features predictive splicing dysregulation in transcriptome-wide datasets. Finally, I examine the evolutionary dynamics of left- and right-handed 5→ SS and propose a model of intron evolution in which codon and intron phase constraints in protein-coding genes shape both minor-to-major intron conversion and transitions between left- and right- 5→ SS subclasses.
Ph.D.
Massachusetts Institute of Technology
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https://rightsstatements.org/page/InC-EDU/1.0/
Quantitative modeling of 5' splice site subclass regulation and evolution
Thesis
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Doctoral
Doctor of Philosophy
Publication
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<Abstract>Pre-mRNA splicing is an essential molecular process required for eukaryotic gene expression. In this thesis, I present a previously unknown mechanism of splicing regulation where a family of splicing factors, the LUC7 family, compete to differentially impact 5→ splice site (5→ SS) selection in a sequence-dependent manner. I quantitatively characterize two major subclasses of 5→ SS in eukaryotes and outline distinctive features of 5→ SS in exons affected by the three human LUC7 paralogs: LUC7L2 and LUC7L enhance splicing of “right-handed” 5→ SS that exhibit stronger consensus matching on the intron side of the nearly-invariant / GU, while LUC7L3 boosts splicing of “left-handed” 5→ SS with stronger consensus matching upstream of the /GU. Using a range of experimental systems, from human cells to mutant plants, I show that LUC7 paralogs have opposing effects on these two 5→ SS subclasses and that this regulatory mechanism likely originated in the last common ancestor of animals and plants over 1.5 billion years ago. I further evaluate a competing model of 5→ SS subclass regulation involving METTL16- mediated U6 snRNA modification and reconcile both models by devising computational tools that identify sequence features predictive splicing dysregulation in transcriptome-wide datasets. Finally, I examine the evolutionary dynamics of left- and right-handed 5→ SS and propose a model of intron evolution in which codon and intron phase constraints in protein-coding genes shape both minor-to-major intron conversion and transitions between left- and right- 5→ SS subclasses.</Abstract>
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