Clara Louise Kielkopf, Ph.D.

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Up to half of human genetic diseases, including cancers and neuromuscular disorders, are associated with defects in splicing of pre-mRNA transcripts. Almost all human genes contain intervening, non-coding sequences that must be spliced from the pre-mRNA transcripts before translation into proteins. The majority of these are alternatively spliced to encode a number of protein variants depending on different cellular needs. My laboratory uses structural and biophysical techniques to understand how pre-mRNA splice sites are recognized in normal cells, and to suggest possible means for treating defective pre-mRNA splicing observed in human genetic diseases.

The major focus of the laboratory is the mechanism of 3' splice site recognition by complexes with the essential pre-mRNA splicing factor, U2 Auxiliary Factor (U2AF). Using biophysical methods including X-ray crystallography, fluorescence anisotropy, calorimetry, and surface plasmon resonance, we characterize the three-dimensional shapes and energetic forces that enable U2AF to recognize the pre-mRNA splice site. We recently determined a series of X-ray structures of U2AF a series of pre-mRNA splice site sequences. An immediate goal of the laboratory is to determine structures of higher order 3' splice site complexes, and the specific roles of phosphorylation and ATP hydrolysis during assembly of these structures. Knowledge of the key interactions between splicing factors and the pre-mRNA will assist the development of chemotherapeutics targeted at the level of pre-mRNA splicing.

Current Appointments

• Assistant Professor - Department of Biochemistry and Biophysics (SMD)

Lab Website

http://dbb.urmc.rochester.edu/labs/kielkopf/

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Recent Journal Articles
Showing the 5 most recent journal articles. (19 available)
Manceau, V.; Kielkopf, C. L.; Sobel, A.; Maucuer, A.;. "Different requirements of the kinase and UHM domains of KIS for its nuclear localization and binding to splicing factors". J Mol Biol 381 (2008): 748-62.
Kumar AO; Swenson MC; Benning MM; Kielkopf CL. "Structure of the central RNA recognition motif of human TIA-1 at 1.95A resolution." Biochemical and biophysical research communications. 2008; 367(4):813-9. Epub 2008 Jan 15.
Lin Y; Kielkopf CL. "X-ray structures of U2 snRNA-branchpoint duplexes containing conserved pseudouridines." Biochemistry. 2008; 47(20):5503-14. Epub 2008 Apr 25.
Jenkins JL; Shen H; Green MR; Kielkopf CL. "Solution conformation and thermodynamic characteristics of RNA binding by the splicing factor U2AF65." The Journal of biological chemistry. 2008; 283(48):33641-9. Epub 2008 Oct 08.
Thickman KR; Sickmier EA; Kielkopf CL. "Alternative conformations at the RNA-binding surface of the N-terminal U2AF(65) RNA recognition motif." Journal of molecular biology. 2007; 366(3):703-10. Epub 2006 Dec 02.