Pre-mRNA Splicing for Treatment of Human Disease
Most human cancers, leukemia’s and many metabolic disorders are accompanied by severe defects in pre-mRNA splicing. RNA splicing is a key step of gene expression and is essential for human life. A major focus of my laboratory is to understand at a molecular level how the splicing machinery identifies sites for excision from gene transcript RNAs, which in turn changes the proteins produced. We have characterized the three-dimensional shapes of human splicing proteins recognizing one another and the gene transcript RNA [for example, Kielkopf et al. (2001); Kielkopf, Luecke, Green (2004); Sickmier et al. (2006); Jenkins et al. (2013); Wang et al. (2013)] (shown above), both at high resolution by X-ray crystallography and in solution by small-angle X-ray scattering and nuclear magnetic resonance. Through this work, we identify a network of interactions responsible for recognizing human splice sites.
Currently, our major goals investigate:
- Determine the three-dimensional shape of the full assembly of human splicing proteins bound to the gene transcript
- Understand the function of splicing factor phosphorylation and its potential to serve as a drug target
- Leverage emerging views of splice site signal recognition as a basis to predictably target and correct defective splice sites
Altogether, this work would elucidate:
- How pre-mRNA splicing fidelity is ensured in humans
- How pre-mRNA splice site recognition is achieved in normal cells and altered in cancers and genetic disease
Current funding includes R01 GM070503 (Kielkopf):
Molecular recognition during pre-mRNA splicing and F32 GM097916 (Laird):
Molecular Means for 3' Splice Site Recognition by U2AF35.
- U2AF1 mutations alter sequence specificity of pre-mRNA binding and splicing.Leukemia. 29, 909-17. (2015 Apr 01).
- Structure-guided U2AF65 variant improves recognition and splicing of a defective pre-mRNA.Proc Natl Acad Sci U S A. 111, 17420-5. (2014 Dec 09).
- Cancer-relevant splicing factor CAPERα engages the essential splicing factor SF3b155 in a specific ternary complex.J Biol Chem. 289, 17325-37. (2014 Jun 20).