The overarching goal of this project is to elucidate the molecular mechanism governing the catalysis and regulation of histone modification enzymes. We will use our Chaperone-Enabled Biology and Structure (CEBS) technology platform to study an important, but challenging group of epigenetic regulating enzymes. The lysine-specific histone methyltransferases (HMT) and their complementary partners, lysine demethylases (KDM) function as key mediators of epigenetic signaling through their actions as "writers and erasers" of post-trahslational modifications on histone proteins. Numerous recent studies have highlighted the importance of lysine methylation of histones leading to direct impact on DNA replication, repair, recombination, gene silencing, imprinting and RNA processes making these enzymes potential key targets for drug development. However, progress in gaining fundamental knowledge about structure-function relationships governing their modes of operation has been slow because they are multidomain proteins and have been recalcitrant to both structural and functional analyses. To overcome the existing barriers, we will generate specialized reagents called "synthetic affinity binders" or sABs that will be used as chaperones for crystallization, as well as customized affinity reagents for cell biologically applications. To accomplish our objectives we have assembled a world-class team of investigators that will exploit sAB reagents for both structure determination and high level biological assays. A major emphasis of our approach is to identify and structurally/biochemically characterize the molecular complexes in which the HMTs and KDMs function by using sABs that stabilize and enhance crystallization of the complexes. Thus, CEBS effort will rely on close ties to the large high throughput centers for protein production and structure determination of individual enzymes in multiple conformational states, enzyme-substrate complexes and multiprotein complexes. A unique strength of our approach is that we will provide crystallization chaperones and the information on interaction partners to the high throughput centers to greatly increase the probability of success of structure determination.
Lysine-specific histone methyltransferases and histone demethylases are critical enzymes in the epigenetic regulation of DNA replication and repair, as well as recombination and gene expression and repression. Mutations in these enzymes have been implicated in a number of diseases. These studies will provide the biological rationale and structural basis for drug development.
|Galilee, Meytal; Britan-Rosich, Elena; Griner, Sarah L et al. (2016) The Preserved HTH-Docking Cleft of HIV-1 Integrase Is Functionally Critical. Structure 24:1936-1946|
|Dominik, Pawel K; Borowska, Marta T; Dalmas, Olivier et al. (2016) Conformational Chaperones for Structural Studies of Membrane Proteins Using Antibody Phage Display with Nanodiscs. Structure 24:300-9|
|Mateja, Agnieszka; Paduch, Marcin; Chang, Hsin-Yang et al. (2015) Protein targeting. Structure of the Get3 targeting factor in complex with its membrane protein cargo. Science 347:1152-5|
|Borowska, Marta T; Dominik, Pawel K; Anghel, S Andrei et al. (2015) A YidC-like Protein in the Archaeal Plasma Membrane. Structure 23:1715-24|
|Nocula-Lugowska, Malgorzata; Lugowski, Mateusz; Salgia, Ravi et al. (2015) Engineering Synthetic Antibody Inhibitors Specific for LD2 or LD4 Motifs of Paxillin. J Mol Biol 427:2532-47|
|Stuwe, Tobias; Correia, Ana R; Lin, Daniel H et al. (2015) Nuclear pores. Architecture of the nuclear pore complex coat. Science 347:1148-52|
|Stuwe, Tobias; Bley, Christopher J; Thierbach, Karsten et al. (2015) Architecture of the fungal nuclear pore inner ring complex. Science 350:56-64|
|Zhong, Nan; Loppnau, Peter; Seitova, Alma et al. (2015) Optimizing Production of Antigens and Fabs in the Context of Generating Recombinant Antibodies to Human Proteins. PLoS One 10:e0139695|
|Mukherjee, Somnath; Ura, Marcin; Hoey, Robert J et al. (2015) A New Versatile Immobilization Tag Based on the Ultra High Affinity and Reversibility of the Calmodulin-Calmodulin Binding Peptide Interaction. J Mol Biol 427:2707-25|
|Dominik, Pawel K; Kossiakoff, Anthony A (2015) Phage display selections for affinity reagents to membrane proteins in nanodiscs. Methods Enzymol 557:219-45|
Showing the most recent 10 out of 25 publications