个人简介

Ph.D.: California Institute of Technology, 1994 Postdoctoral Fellow: National Science Foundation Postdoctoral Fellow, Scripps Research Institute, 1994-96

研究领域

Molecular Biophysics Nucleic Acids Proteins & Enzymology Structural Biology Biophysics

Telomere Biology Telomeres are specialized nucleoprotein structures at the ends of eukaryotic chromosomes that are essential for chromosome stability and cellular proliferation. Telomeric DNA does not encode for proteins, instead it consists of tandem repeats of TG-rich sequences of double-stranded DNA that terminate in a 3¢ single-stranded DNA overhang. Protection of this overhang is essential. When left unprotected, this overhang initiates DNA damage responses that lead to catastrophic events permanently damaging the genome and resulting in apoptosis or senescence. Furthermore, telomere shortening due to the inability of the DNA-replication machinery to fully replicate the ends is a critical mechanism of tumor suppression as well as a hallmark of aging. Continually proliferating cells maintain adequate telomeres through the action of the reverse transcriptase telomerase.Telomeres are important to human health because dysregulation of either telomere protection or telomerase activity causes many human diseases. Notably, over 90% of human cancers activate telomerase for continued proliferation. Our research in this area aims to understand how telomere-associated proteins protect and maintain telomeres. Key questions include how subunits of the telomerase enzyme contribute to activity, how the single-strand DNA overhang is shielded from the DNA-damage machinery, and whether capping activity also regulates telomerase action. We develop this knowledge by first understanding the core activities of key telomere factors, then testing these activities in a reconstituted telomerase assay and validating our knowledge directly in the organism. Plasticity in Molecular Recognition Many biologically critical recognition events involve the specific binding of flexible ligands such as single-stranded (ss) DNA, RNA, peptides and carbohydrates. Structural plasticity, defined as the ability of an interface to adopt alternate conformations when bound to different ligands, has been invoked to explain binding specificity and promiscuity in several protein/ligand systems. Furthermore, an understanding of the malleability of a binding interface is increasingly recognized as key to predicting its binding activity and specificity. Discerning the scope and mechanisms of rearrangements at binding interfaces is essential to understanding the biophysics of molecular recognition events. The focus of this proposal is to investigate the extent of structural plasticity in the recognition of these flexible ligands. We use the recognition of ssDNA by the telomere end-binding proteins as the predominant model to characterize the contribution of structural plasticity to recognition. The telomere-end binding proteins Pot1 and Cdc13 bind the conserved 3’ ssDNA overhang at telomeres. This binding is required for cellular viability. However, the sequence of the overhang is somewhat variable, meaning that these proteins need to bind divergent ligands while maintaining exquisite specificity. Extensive evidence suggests that the protein/nucleic acid interface adopts altered configurations in the presence of different ligands that bind with similar affinities. We are investigating the hypothesis that this structural plasticity is important for specificity. Moreover, the malleability of the interface may further contribute to function by providing a way to physically alter the structure and accessibility of the 3’ end. We us an integrated set of strategies to address this question, ranging from determination of high-resolution structures to in vivo assessment of activities.

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K. A. Lewis, D. A. Pfaff, J. N. Earley, S. E. Altschuler, and D. S. Wuttke, “The Tenacious Recognition of Yeast Telomere Sequence by Cdc13 is Fully Exerted by a Single OB-Fold Domain,” Nucleic Acids Res., 2013, Sept 20 epub ahead of print T. H. Dickey, S. E. Altschuler and D. S. Wuttke, “-Single-stranded DNA-binding Proteins: Multiple Domains for Multiple Functions,” Structure, 2013, 21, 1074-84 T. H. Dickey, M. A. McKercher, and D. S. Wuttke, “Nonspecific Recognition Is Achieved in Pot1pC through the Use of Multiple Binding Modes,” Structure, 2013, 21, 121-32 S. E. Altschuler, J. E. Croy, and D. S. Wuttke, “A Small Molecule Inhibitor of the S. pombe Pot1 binding to telomeric DNA,” Biochemistry, 2012, 51, 7833-45 K. A. Lewis and D. S. Wuttke, “Telomerase and Telomere-Associated Proteins: Structural insights into mechanism and evolution,” Structure, 2012, 20, 28 - 39 S. E. Altschuler, T. H. Dickey, and D. S. Wuttke, “Schizosaccharomyces pombe Protection of Telomeres 1 Utilizes Alternate Binding Modes To Accommodate Different Telomeric Sequences” Biochemistry, 2011, 50, 7503-15 E. K. Mandell, A. D. Gelinas, D. S. Wuttke, V. Lundblad, “Sequence-specific binding to telomeric DNA is not a conserved property of the Cdc13 DNA binding domain,”. Biochemistry, 2011, 50, 6289-91 J. Lee, E. K. Mandell, T. Rao, D. S. Wuttke and V. Lundblad, “Investigating the role of the Est3 protein in yeast telomere replication,” Nucleic Acids Res., 2010, 38, 2279-2290 A. D. Gelinas, M. Paschini, F. E. Reyes, A. Héroux, R. T. Batey, V. Lundblad and D. S. Wuttke, “Telomere capping proteins are structurally related to RPA with an additional telomere-specific domain” Proc. Natl. Acad. Sci. USA, 2009, 106, 19298-19303 J. E. Croy and D. S. Wuttke, “Insights into the dynamics of specific telomeric single-stranded DNA recognition by Pot1pN,” J. Mol. Biol., 2009, 387, 935-938 J. E. Croy, S. E. Altschuler, N. E. Grimm, and D. S. Wuttke, “Nonadditivity in the recognition of single-stranded DNA by the Schizosaccharomyces pombe protection of telomeres 1 DNA-binding domain, Pot1-DBD,” Biochemistry, 2009, 48, 6864-6875 D. C. Zappulla, J. N. Roberts, K. Goodrich, T. R. Cech, and D. S. Wuttke, “Inhibition of yeast telomerase action by the telomeric ssDNA-binding protein, Cdc13p,” Nucleic Acid Res., 2009, 37, 354-367 J. E. Croy, J. L. Fast, N. E. Grimm, and D. S. Wuttke, “Thermodynamic characterization of specific single-stranded telomeric DNA binding to the S. pombe, Protection of telomeres 1 (POT1), protein, Biochemistry, 2008, 47, 4345-4358 A. E. Eldridge and D. S. Wuttke, “The mechanism of recognition of ssDNA by the Cdc13-DBD,” Nucleic Acid Res., 2008, 36, 1624-33 D. L. Theobald and D. S. Wuttke, “Accurate structural correlations from maximum likelihood superpositions,” PLOS Comp. Biology, 2008, e43 J. R. Croy and D. S. Wuttke, “Themes in ssDNA recognition by telomere end-protection proteins,” TIBS, 2006, 31, 516-525 D. L. Theobald and D. S. Wuttke, THESEUS: Maximum likelihood superpositioning and analysis of macromolecular structures, Bioinformatics, 2006, 22, 2171-2 A. M. Eldridge, W. A. Halsey, and D. S Wuttke, “Identification of the determinants for the specific recognition of single-strand telomeric DNA by Cdc13,” Biochemistry, 2005, 45, 871-879 D. L. Theobald and D. S. Wuttke, “Divergent evolution within protein superfolds inferred from profile-based phylogenetics” J. Mol. Biol., 2005, 354, 722-737 D. L. Theobald and D. S. Wuttke, “Prediction of multiple tandem OB-folds in telomere end-binding proteins Pot1 and Cdc13” Structure, 2004, 12, 1877-1879. R. M. Mitton-Fry, E. M. Anderson, L. W. Glustrom, D. L. Theobald and D. S. Wuttke, “Structural basis for telomeric single-stranded DNA recognition by yeast Cdc13,” J. Mol. Biol., 2004, 338, 241-255. Daniel M. Strauss, Leslie W. Glustrom, and Deborah S. Wuttke. "Towards an understanding of the poliovirus replication complex: the solution structure of the soluble domain of the poliovirus 3A protein," J. Mol. Biol., 330, 225-234 (2003). Douglas L. Theobald, Rachel M. Mitton-Fry, and Deborah S. Wuttke. "Nucleic acid recognition by OB-fold proteins," Annu. Rev. Biophys. and Biomol. Struct., Feb. 18 (2003). Emily M. Anderson, and Deborah S. Wuttke. "Site-directed mutagenesis reveals the thermodynamic requirements for single-stranded DNA recognition by the telomere-binding protein Cdc13," Biochemistry, 42, 3751-3758 (2003). Deanna Dahlke Ojennus, Sarah E. Lehto, and Deborah S. Wuttke. "Electrostatic interactions in the reconstitution of an SH2 domain from constituent peptide fragments," Protein Science, 12, 44-55 (2003).