mspies@life.illinois.edu
493 Roger Adams Lab
Office: (217) 244-9493
Fax: (217) 244-5858
Mail to:
Department of Biochemistry
University of Illinois
600 S Mathews Ave
Urbana, IL 61801
Maria Spies
Assistant Professor of Biochemistry and Biophysics
Education
B.S. 1994 St. Petersburg State Polytechnic University, St. Petersburg, Russia
M.S. 1996 St. Petersburg State Polytechnic University, St. Petersburg, Russia
Ph.D. 2000 Osaka University, Osaka, Japan
Postdoc. 2000-2005 University of California, Davis
Teaching Interests
Molecular mechanisms of DNA helicases and DNA motor proteins; mechanistic aspects of protein-nucleic acids and protein-protein interactions; homologous genetic recombination; DNA repair; molecular motors
Our work focuses on the genome caretakers involved in homologous recombination and nucleotide excision repair. The following systems are being currently pursued in my lab:
Motor components of the DNA repair machineries, in particular DNA helicases XPD, Bach1, Fbh1. Central to many DNA-repair machineries are DNA helicases, ubiquitous molecular motors that use the energy of nucleoside triphosphate (NTP) hydrolysis to move directionally along nucleic acids. Translocation by helicases is coupled to other thermodynamically unfavorable processes including separation of nucleic acid duplexes and disassembly of protein-nucleic acid complexes. The question we try to answer is how these structurally similar enzymes perform an amazingly diverse set of activities, how they utilize unique structural features incorporated into otherwise conserved motor cores and how other players in the genome maintenance pathways modulate activities of selected helicases adapting them to desired cellular tasks.
Recombination mediators: Lesions affecting both strands of the DNA duplex are constantly generated in the cell and have to be mended to prevent genomic instability leading to cancer or cellular senescence. Most accurately this is achieved through the process of homologous genetic recombination, which depends on assembly of the Rad51 recombinase into continuous nucleoprotein filament on ssDNA generated at the site of damage. Rad51 nucleoprotein filament formation is opposed by ssDNA sequestering by ssDNA binding proteins and is highly controlled by a group of recombination mediator proteins. We study one of these recombination mediators, Rad52 protein and its effect of Rad51 activities.
APPROACHES:
We study the DNA repair processes at the most fundamental level by first deconstructing the macromolecular ensembles orchestrating the DNA repair events down to the level of individual proteins. By combining classical and single-molecule biochemistry for analysis of the key players in these DNA repair pathways, we then investigate their molecular mechanism and how other protein partners affect their action. These studies allow us to gain fundamental biochemical knowledge of accurate DNA repair and help us to identify features of the genome caretaker proteins that can be exploited in designing the new therapeutics.
Representative Publications
Honda, M., Park, J., Pugh, R. A., Ha, T., and Spies, M. Single-molecule analysis reveals differential effect of ssDNA binding proteins on DNA translocation by XPD helicase: (2009) Submitted to Mol.Cell
Grimme, J. M., Wright, R., Rothenberg, E., Mazin, A., Ha, T., and Spies, M., Interplay between DNA binding and annealing modes in human Rad52 recombination mediator protein (2009) Submitted to EMBO J.
Rothenberg, E., Grimme, J. M., Spies, M. and Ha, T.,Rad52 protein mediates directionally biased homology search and DNA annealing through continuous association of two Rad52-ssDNA complexes. (2008) PNAS 105 (51) 20274-20279
Pugh, R. A., Lin, Y., Eller C., Leesley H., Cann, I. K.O., and Spies, M. Ferroplasma acidarmanus RPA2 facilitates efficient unwinding of forked DNA substrates by monomers of XPD helicase. (2008) JMB 383(5): 982-98
Lin, Y., Lin, L.-J., Sriratana, P., Coleman, K., Ha, T., Spies, M., Cann, I.K.O. Engineering of functional replication protein A homologs based on insights in evolution 1 of 2 oligonucleotide/oligosaccharide binding (OB) folds. (2008); J. Bac, 190 (17) 5766-5780
Pugh, R.A., Honda, M., Leesley, H., Thomas, A., Lin, Y., Nilges, M.J., Cann, I.K.O., Spies, M. The iron-containing domain is essential in RAD3 helicases for coupling of ATP hydrolysis to DNA translocation and for targeting the helicase to the ssDNA-dsDNA junction. (2008) JBC, 283(3):1732-43.;
Pugh, R. A., and Spies, M., "DNA Helicases, Chemistry and Mechanisms of" (in Wiley Encyclopedia of Chemical Biology) (2008) in press
Spies, M., Amitani, I., Baskin, R.J., and Kowalczykowski, S. C. Translocation by single-molecules of a RecBCD motor mutant reveals a switch in motor subunit usage at χ. (2007) Cell, 131: 694-705;
Grimme, J. M., and Spies, M., "DNA helicases in homologous recombination" (in "Molecilar Genetics, Biophysics and Medicine Today" (Bresler Memorial Lectures –II) Editor: Vladislav Lanzov), PNPI Press, 2007, St. Petersburg, Russia ISBN 5-86763-197-4) 123-140
Spies, M., and Kowalczykowski, S. C., The RecA-binding locus of RecBCD enzyme defines a generalized domain for recruitment of DNA strand exchange proteins. (2006) Molecular Cell, 21: 573-580.; (Faculty of 1000 recommended article)
Spies, M., Dillingham, M.S., and Kowalczykowski, S. C., "Translocation by the RecB motor is an absolute requirement for Χ-recognition and RecA protein loading by RecBCD enzyme". (2005) JBC, 280: 37078-37087;
Spies, M., and Kowalczykowski, S.C., "Homologous recombination by RecBCD and RecF pathways" in "The Bacterial Chromosome". (ASM Press, Washington, D.C., 2004), 389-403
Spies, M., Bianco, P.R., Dillingham, M.S., Handa, N., Baskin, R.J., and Kowalczykowski, S.C., A molecular throttle: The recombination hotspot, χ controls DNA translocation by the RecBCD helicase. (2003) Cell, 114: 647-654.; (Featured Article; News & Features at Bio.com)
Dillingham, M.S., Spies, M., and Kowalczykowski, S.C., RecBCD enzyme is a bipolar DNA helicase. (2003) Nature, 423(6942): 893-897; (News & Views in Nature Structural Biology 10, 499 - 500 (2003); Highlight in Nature Reviews Molecular Cell Biology 4, 513 (2003); Faculty of 1000 must read article
Spies, M., Dillingham, M.S., and Kowalczykowski, S.C., "DNA helicases" in McGraw-Hill 2003 Yearbook of Science & Technology. (McGraw-Hill Professional, 2003), 95-98
Spies, M., Kil, Y., Masui, R., Kato, R., Kujo, C., Oshima, T., Kuramitsu, S., and Lanzov, V., The RadA protein from a hyperthermophilic archaeon Pyrobaculum islandicum is a DNA-dependent ATPase that exhibits two disparate catalytic modes, with a transition temperature at 75°C. (2000) The European Journal of Biochemistry, 267(4): 1125-1137.