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High-Resolution Complexome Profiling by Cryoslicing BN-MS Analysis
Published on: October 15, 2019
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Spliceosome Profiling Visualizes Operations of a Dynamic RNP at Nucleotide Resolution
Jordan E Burke1, Adam D Longhurst1, Daria Merkurjev2
1Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94158, USA.
Cell
|May 5, 2018
Summary
New tools globally profile spliceosome-bound RNA in vivo, revealing splicing regulation and coevolution across yeast species. This quantitative method advances understanding of this essential RNP complex in gene expression.
Area of Science:
- Molecular Biology
- Genomics
- Evolutionary Biology
Background:
- Understanding spliceosome function in vivo has lagged behind structural biology advances.
- The spliceosome is a crucial ribonucleoprotein (RNP) complex in eukaryotic gene expression.
Purpose of the Study:
- To develop and apply novel methods for globally profiling spliceosome-bound RNA at nucleotide resolution.
- To investigate spliceosome dynamics, substrate selection, and coevolution across diverse yeast species.
Main Methods:
- Development of a global spliceosome profiling technique.
- Application to three yeast species representing significant evolutionary divergence.
- Utilizing statistical modeling to analyze RNA processing events.
Main Results:
- Detection of canonical and non-canonical splicing events (interrupted, recursive, nested).
- Identification of intron size, position, and number as key factors in substrate progression.
- Discovery of ATP-dependent substrate discard post-spliceosome assembly, linked to intron retention.
Conclusions:
- Spliceosome profiling is a generalizable, quantitative technology for studying RNP complexes.
- Revealed insights into spliceosome-transcriptome coevolution and splicing regulation.
- Advanced the understanding of spliceosome dynamics and its role in gene expression.
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